Ligand/receptor specificity exchangers delivering antibodies to receptors on pathogen

FIELD: medicine, biotechnology, pharmacy.

SUBSTANCE: invention relates to exchangers of ligand/receptor specificity delivering antibodies to receptors on pathogen. In particular, invention describes variants related to manufacturing and using exchangers of ligand/receptor specificity. Exchangers comprise at least one specificity domain containing ligand for receptor wherein ligand is not antibody or its part, and at least one antigenic domain combined with abovementioned specificity domain wherein antigenic domain comprises epitope of pathogen or toxin. Advantage of the invention involves enhanced specificity in delivery of drug.

EFFECT: improved and valuable properties of exchangers.

30 cl, 5 tbl, 6 ex

 

The scope of the invention

The present invention relates generally to compositions and methods for prevention and treatment of human diseases, including, but not right restrictions, such pathogens as bacteria, yeast, parasites, fungus, viruses and cancer. In particular, the described embodiments of the relate to the production and use of obmenivatsa specificity of ligand/receptor that redirect existing entity antibodies to receptors that are present on the pathogen.

The existing level of technology

Infections caused by such pathogens as bacteria, yeast, parasites, fungi, as well as the emergence and spread of cancer, pose serious problems for the health of all animals, including humans, livestock and Pets. This health hazard is enhanced by the emergence of strains resistant to vaccination and/or treatment. In the past, practitioners pharmacists relied on traditional methods of drug development to develop safe and effective compounds for the treatment of these diseases. Traditional methods of drug development generally contained a blind test molecules candidate potential drug, often selected at random, in the hope that any potential medication can be an effective + treatment the drug for a certain disease. However, with the advent of molecular biology the focus of drug development has shifted to the identification of molecular targets associated with etiological agent, and to the design of compounds that interact with these molecular targets.

One promising class of molecular targets are receptors located on the surface of bacteria, yeast, parasites, fungi, viruses and cancer cells, especially those receptors that provide the attachment to the host cell or to a protein of the host (for example, extracellular matrix protein). Research in this area primarily focus on the identification of the receptor and its ligand and on the detection of molecules that disrupt the interaction of the ligand with the receptor, thereby blocking the adhesion to the cage or squirrel master.

For example, many pathogenic bacteria (for example, Staphylococcus aureus) produce adhesion receptors (e.g., ClfA, Efb and FnBPA), which are able to bind with extracellular matrix proteins of the host (e.g., fibrinogen, fibronectin and laminin). (Flock, Mol. Med. Today 5:532-533 (1999)). Researchers have shown that the commitment of some bacteria to extracellular matrix proteins of the host may be blocked using peptides corresponding to portions of the extracellular matrix protein of the host. (Pei et al., Infection and Immunity 67():4525-4530 (1999)). Similarly, many viruses have receptors that interact with proteins present on the surface of host cells. (See, for example, U.S. patent No. 5942606 and 5929220). Researchers have shown that a fragment of a T4 glycoprotein (a protein of the host cell) may interact with gp120 of human immunodeficiency virus (HIV), and peptide T4 can be used to prevent or treat HIV infection. (See, for example, U.S. patent No. 6093539). Additionally, many types of cancer cells Express receptors that interact with extracellular matrix proteins of the host, and researchers have shown that molecules that block integrin receptors, can be used for inhibiting the accession of tissue, metastasis, angiogenesis and tumor growth. (See, for example, U.S. patent No. 6066648, 6087330, 5846536, 5766591 and 5627263). Although these inhibiting peptides have promising therapeutic potential, there is still a need for new compositions and methods for treating and preventing infections caused by pathogens and other diseases.

The invention

This invention relates to the production, characterization and use of new agents that bind receptors on the pathogens and redirects antibodies available subject to the pathogen. The options for implementation include obmenival specificity is Iganga/receptor, having at least one domain specificity, containing the ligand for the receptor, and at least one antigenic domain, coupled with the aforementioned domain specificity, and mentioned antigenic domain contains the epitope of a pathogen or toxin.

Some embodiments of obmenivalas specificity of ligand/receptor have domain specificity, containing at least three consecutive amino acids of a peptide selected from the group consisting of extracellular matrix protein, a ligand for receptors on the virus and the ligand for a receptor on a cancer cell. In some aspects of this version of the runtime, for example, the peptide is an extracellular matrix protein selected from the group consisting of fibrinogen, collagen, vitronectin, laminin, plasminogen, thrombospondin and fibronectin. Preferably, the extracellular matrix protein comprises at least 3 amino acids of the alpha-chain of fibrinogen, and in the most preferred versions of the ligand contains a sequence arginine-glycine-AC partat (RGD).

In other embodiments, the above peptide is a ligand for a receptor on a virus selected from the group consisting of a T4 glycoprotein and protein shell of the virus of hepatitis B. In other aspects of this option, perform the peptide is a ligand for the recipe of the RA on the cancer cell, selected from the group consisting of a ligand for HER-2/neu and ligand for integranova receptor. Preferred embodiments of the contain the domain specificity, containing the sequence provided by one of the ID. PEFC. No. 1-42.

Obmanivali specificity of ligand/receptor described here, interact with the receptor on the pathogen. In some embodiments, execution of the receptor is bacterial adhesion receptor, such as bacterial adhesion receptor selected from the group consisting of a binding protein of the extracellular fibrinogen (Efb), binding protein collagen, protein, vitronectin binding protein, laminin binding protein, plasminogen binding, protein binding thrombospondin, aggregating factor A (ClfA), aggregating factor B (ClfB), a protein that binds fibronectin, coagulase and protein extracellular adhesion.

Obmanivali specificity of ligand/receptor described here also interact with the antibody present in the subject. In some embodiments execute, for example, antigenic domain contains at least three amino acid peptide selected from the group consisting of a protein of herpes simplex virus, the protein of hepatitis B protein of TT virus and protein of poliovirus. In desirable embodiments perform obmenival special is licnosti ligand/receptor contains antigenic domain, which is the protein of herpes simplex virus containing a sequence selected from the group consisting of ID. PEFC. No. 53 and ID. PEFC. No. 54. In other desirable embodiments, execution of the antigenic domain is a protein of hepatitis B-containing sequence, provide one of the ID. PEFC. No. 49, ID. PEFC. No. 50, ID. PEFC. No. 52 and ID. PEFC. No. 59.

Some obmanivali specificity of ligand/receptor also contain antigenic domain, which is a protein of TT virus containing the sequence provided by one of the ID. PEFC. No. 43-47 and ID. PEFC. No. 55-58. Obmanivali specificity of ligand/receptor may also be antagony domain, which is a protein of poliovirus containing a sequence selected from the group consisting of ID. PEFC. No. 48 and ID. PEFC. No. 51. Preferably, obmenival specificity of ligand/receptor contains an antigenic domain that interacts with the antibody with high titer. In some embodiments execute, for example, antigenic domain specifically binds to an antibody in animal serum which was diluted to approximately 1:100-1:1000 or more. Obmanivali specificity ID. PEFC. No. 60-105 are variants of execution of the invention.

Aspects of the invention also relate to methods of treating or preventing infection or proliferation p is tohana. For example, one approach includes a method of treating and preventing bacterial infections. This method is carried out by providing the subject a therapeutically effective amount of obmenivalas specificity of ligand/receptor, and mentioned obmenival specificity of ligand/receptor contains the domain specificity with a ligand that interacts with a receptor on the bacteria and antigenic domain that contains the epitope for the pathogen or toxin. Also a variant of execution is a method of treating or preventing a viral infection. Accordingly, a method of treating or preventing a viral infection is carried out by supplying the subject a therapeutically effective amount of obmenivalas specificity of ligand/receptor, and mentioned obmenival specificity of ligand/receptor contains the domain specificity containing a ligand that interacts with receptors on the virus and antigenic domain that contains the epitope for the pathogen or toxin. Similarly, variant execution is a method of treatment or prevention of cancer, and this method can be performed by supplying the subject a therapeutically effective amount of obmenivalas specificity of ligand/receptor, and mentioned obmenival specificity of ligand/receptor contains a domain specific the STI, containing a ligand that interacts with a receptor on the cancer cell, and antigenic domain that contains the epitope for the pathogen or toxin.

Detailed description of the invention

The following describes the production, characterization and use of new agents that bind receptors on the pathogens and redirect antibodies available subject to the pathogen. The term "obmenival specificity of antigen/antibodies known in the art and refers to a molecule containing the amino acid sequence corresponding to the amino acid sequence of the antibody (e.g., plot, defining complementarity)associated with the amino acid sequence which binds to a specific antibody (e.g., epitope of a pathogen). (See, for example, Sällberg et al., Biochemical &Biophysical Research Communications, 205:1386-90 (1994) and U.S. patent No. 5869232 and 6040137). Obmanivali specificity of antigen/antibody can redirect antibodies available subject to the pathogen, and these exchanges agents have therapeutic and diagnostic use. (Ibid.).

As described here perform belong to the second generation, trading agents, called "obmanivali specificity of ligand/receptor". Unlike obmenivatsa specificity of antigen/antibody, obmanivali specificity of ligand/Retz is ptor not contain sequences, found in the antibody. Instead, obmanivali specificity of ligand/receptor contain the first domain contains a ligand for a receptor, and a second domain that contains the epitope of a pathogen or toxin. Thus, for the purposes of this disclosure, the term "obmanivali specificity of ligand/receptor" refers to exchanging agent containing "domain specificity", having at least one ligand for a receptor ("ligand" is not an antibody or part of it), coupled with "antigenic domain having at least one epitope of a pathogen or toxin (e.g., pertussis toxin or cholera toxin).

Obmanivali specificity of ligand/receptor may contain more than domain specificity and antigenic domain. For example, some obmanivali specificity of ligand/receptor contain many domain specificity and/or antigenic domains. Obmanivali specificity of ligand/receptor with multiple domain-specific and/or more antigenic domains, called "multipersonality"because more than one domain specificity and/or antigenic domain, combined in tandem. Other embodiments of the are obmenivalas specificity of ligand/receptor, which contain, in addition to domain specificity and antigenic domain sequence that facilitates PTS is STCU (for example, polyhistidine tail), linkers (e.g., Biotin and/or avidin, or streptavidin, or flexible appendages phage 8 (8-linkers)) and of a sequence or modification, which either provide stability obmenivalas specificity of ligand/receptor (e.g., modification, providing resistance to absorption by the protease), or help the decomposition of obmenivalas specificity of ligand/receptor (e.g., sites proteasome splitting). Although the domain specificity and antigenic domain are preferably peptides, some obmanivali specificity of ligand/receptor contain the domain specificity and antigenic domain is made of modified or derived peptides, peptidomimetics, or chemicals.

Variety of obmenivatsa specificity of ligand/receptor is large, as described here perform can communicate with many different receptors on many different pathogens. Thus, the term "pathogen" is used here in a General sense, denoting the etiological agent of the disease in animals, including, but not right restrictions, bacteria, parasites, fungus, mold, viruses and cancer cells. Similarly, the term "receptor" is used in a General sense, denoting a molecule (usually a peptide that differs from the sequence found in the antibody, is about can be a hydrocarbon, the lipid or nucleic acid), interacting with the "ligand" (usually a peptide that differs from the sequence found in the antibody, or a hydrocarbon, lipid, nucleic acid, or a combination). "Receptor"used here is not necessarily subjected to signal transduction and may be involved in a variety of molecular interactions, including, but not right restrictions, adhesion (e.g., integrins) and molecular signaling (e.g., receptors, growth factor). For example, a desirable domain specificity contain the ligand peptide having the sequence represented in the extracellular matrix protein (e.g., fibrinogen, collagen, vitronectin, laminin, plasminogen, thrombospondin and fibronectin), and some domains of the specificity contain a ligand that interacts with bacterial adhesion receptor (e.g., extracellular protein that binds fibrinogen (Efb), a protein that binds collagen, a protein that binds vitronectin, protein, laminin binding, protein binding plasminogen, protein binding thrombospondin, aggregation factor A (ClfA), an aggregating factor B (ClfB), a protein, binding fibronectin, coagulase and protein extracellular adhesion).

In other versions of the domain specificity contains the ligand, have the second peptide sequence, which interacts with the viral receptor (e.g., fragment of a T4 glycoprotein that binds gp120, or a fragment of the preS domain that binds gp170 collection hepadnavirus). In other versions of the domain specificity contains a ligand that interacts with a receptor on a cancer cell (e.g., HER-2/neu (C-erbB2)or such integranova receptor, as vitro-actinomy receptor, leninova receptor, fibronectin receptor, collagen receptor, fibrinogenesis receptor, receptor α4β1, receptor α6β1, receptor α3β1, receptor α5β1, and receptor αvβ3. However, in preferred versions of the domain specificity contains at least 8 amino acids of the alpha-chain of fibrinogen and/or sequence arginine-glycine-aspartic acid (RGD), and the most preferred embodiments of the have domain specificity containing a sequence selected from the group consisting of ID. PEFC. No. 60-105.

Desirable antigenic domains contain the epitope that is recognized by the antibody, which already exists in the subject. For example, many people are immunized against childhood diseases, including, but not right restrictions, smallpox, measles, mumps, rubella and polio. Thus, antibodies to epitopes of these pathogens can barabati Atsa immunized person. Desirable antigenic domains contain the epitope which is found in these aetiological agents.

In some embodiments, execution of antigenic domains interact with the antibody, which was introduced to the subject. For example, the antibody that interacts with antigenic domain on obmanivali specificity of ligand/receptor, may be injected together with obmenivalas specificity of ligand/receptor. Further, the antibody that interacts with obmenivalas specificity of ligand/receptor, in a usual condition does not exist in the subject, but the subject got this antibody by introducing a biological material (for example, serum, blood or tissue). For example, subjects exposed to blood transfusion, receive numerous antibodies, some of which can interact with the antigenic domain of obmenivalas specificity of ligand/receptor.

The most desirable antigenic domains contain the epitope that is recognized by antibody with high titer. By "antibody with high titer" refers to an antibody having a high affinity for the antigen (e.g., epitope in antigenic domain). For example, in fermentopathia immunosorbent analysis (ELISA) antibody with high titer corresponds to the antibody present in the serum sample, which remains positive in the analysis of pokeronline serum to a range of approximately 1:100-1:1000 in a suitable dilution buffer, preferably up to about 1:500. However, the preferred antigenic domains contain the epitope detected in the gG2 protein of herpes simplex virus, s-antigen of hepatitis B (HbsAg), e antigen hepatitis virus (HBeAg), c-antigen of hepatitis B (HBcAg), TT virus and the polio virus or combinations thereof, or contain a sequence selected from the group consisting of ID. PEFC. No. 43-59.

Described here obmanivali specificity of ligand/receptor can be accomplished by generally accepted in the recombinant engineering and/or peptide chemistry techniques. In some embodiments, performing the domain specificity and antigenic domains are performed separately, and then connected to each other (e.g., via linkers, or by binding to a common carrier molecule). In other versions of the domain specificity and antigenic domain are performed as parts of the same molecule. In accordance with one of the approaches obmenival specificity of ligand/receptor, having a domain specificity associated with antigenic domain is produced by using a peptide synthesizer. In accordance with another approach nucleic acid encoding a domain specificity, joined with the antigenic domain is cloned in the expression construct, transfairusa in cells, and obmenival specificity of ligand/is eceptor purified or isolated from cells or the emerged part of the cell.

When obmenival specificity of ligand/receptor obtained, it can be examined to determine its ability to interact with the receptor on the pathogen and/or antibody specific for the antigenic domain. Thus, the term "characteristic analysis" is used to describe the experiment or assess the ability of obmenivalas specificity of ligand/receptor to interact with the receptor on the pathogen or cancer cell or a fragment and/or an antibody specific for the antigenic domain. Some of the characteristic tests, for example, assess the ability of obmenivalas specificity of ligand/receptor contact with the base containing the receptor of the pathogen or its fragment, located on her, or Vice versa. Other characteristic tests evaluate the ability of obmenivalas specificity of ligand/receptor contacted with the antibody specific for the antigenic domain of obmenivalas specificity of ligand/receptor. Other characteristic tests evaluate the ability of obmenivalas specificity of ligand/receptor to influence infection by a pathogen or proliferation of cancer cells in cultured cell lines or diseased animals.

Described here obmanivali specificity of ligand/receptor can be used as active ingredient the RC medicines for the treatment and prevention of pathogenic infection, and cancer in animals, including humans. Pharmaceutical options execution can be formulated in many ways and can contain fillers, binders, emulsifiers, carriers, and other auxiliary agents in addition to obmenival specificity of ligand/receptor. Drugs, containing obmenival specificity of ligand/receptor, can be entered in various ways, including, but not right restrictions, topically, transdermally, parenterally, a gastroenteric path, responcible and resolvename. Obmanivali specificity of ligand/receptor can also be used as coatings for medical equipment and prostheses to prevent infection or the spread of the disease. The number of obmenivalas specificity of ligand/receptor provided in the form of medicine in therapeutic Protocol or applied to the medical device varies depending on the intended use, patient and frequency of administration.

Some of the disclosed methods relate to the introduction of obmenivalas specificity of ligand/receptor to a subject in need of treatment and/or prevention of bacterial infection, fungal infection, viral infection and cancer. In accordance with one approach, to a subject suffering from a bacterial infection, impose on Mediatel specificity of ligand/receptor, containing the domain specificity, interacting with bacterial receptor. Similarly to a subject suffering from a viral infection, can enter obmenival specificity of ligand/receptor, containing the domain specificity of interacting with the viral receptor, and to a subject suffering from cancer, enter obmenival specificity of ligand/receptor, containing the domain specificity of interacting with a receptor on cancer cells. When a complex of the receptor/obmenival specificity, it is assumed that the pathogen or cancer cell excreted by the complement fixation and/or destruction by macrophages.

Are provided methods of treating and preventing diseases such as bacterial, fungal and viral infections, and cancer), in which a subject suffering from a disease, or a subject at risk is identified and then provided a therapeutically effective amount of obmenivalas specificity of ligand/receptor, which interacts with a receptor present on the etiological agent. Accordingly, subjects suffering from a bacterial infection, fungal infection, viral infection or cancer, are identified using standard clinical and diagnostic examination and supplied therapeutically effective the active number of obmenivalas specificity of ligand/receptor, which interacts with a specific pathogen or cancer cell. Although described here obmanivali specificity of ligand/receptor can be administered to all animals at risk in the preventive purposes, it may be desirable to introduce obmanivali specificity of ligand/receptor only to those individuals who belong to the category of high risk (e.g., children, the elderly, and those who have close contact with pathogens). As noted above, individuals with high risk are identified using currently available clinical and diagnostic technologies.

The following section describes in detail the different types of obmenivatsa specificity of ligand/receptor that interact with receptors on the bacteria, parasites, fungi, mold, viruses and cancer cells.

Obmanivali specificity of ligand/receptor that interacts with the receptor on the pathogen

Obmanivali specificity of ligand/receptor that interacts with the receptor on the pathogen, have different chemical structures, but in General, they are characterized by the fact that they contain at least one site that binds to a receptor (domain specificity), and at least one site that interacts with an antibody specific for an epitope of a pathogen or toxin (antigenic domain). P is edocfile obmanivali specificity of ligand/receptor are peptides, but some embodiments of the contain derivatives or modified peptides or coworkers peptide structure. For example, typical based on peptide obmenival specificity of ligand/receptor may be modified so as to have alternates that are not commonly found in the peptide, or may have substituents, which are usually located in the peptide, but implemented in areas that are not normal. In this sense, based on the peptide obmenival specificity of ligand/receptor may be acetiminophen, allyawan or minioven, and the substituents that can be introduced into the peptide for its modifications are not in order restrictions, H, alkyl, aryl, alkenyl, quinil, aromatic compounds, ether, ester, unsubstituted or substituted amine, amide, halogen or unsubstituted or substituted sulfonyl or 5 - or 6-membered aliphatic or aromatic ring. Thus, the term "obmenival specificity of ligand/receptor" is broad and encompasses modified or unmodified peptide patterns, as well as peptidomimetics and chemical structure.

There are many ways of making peptidomimetic that reminds based on peptide obmenival specificity of ligand/receptor. All naturally occurring amino acids, COI is Lithuania in biological production of peptides have L-configuration. Synthetic peptides can be prepared using conventional synthesis methods, using L-amino acids, D-amino acids, or various combinations of amino acids of these two different configurations. Synthetic compounds mimicking the structure and the desired characteristics of the peptide, but does not have undesirable characteristics, such as flexibility (loss of structure) and unlink, known as "peptidomimetics". (See, for example, A.F. Spatola Chemistry and Biochemistry of Amino Acids. Peptides and Proteins (Weistein, C., ed.), Vol.7, pp.267-357, Marcel Dekker, New York (1983), which describes the use of mutilatio of bioisostere [CH2S] as an amide replacement in enkephalinase analogues; and Szelke et al., In peptides: Structure and Function, Proceedings of the Eighth American Peptide Symposium, (Hruby and Rich, Eds.): pp.579-582, Pierce Chemical Co., Rockford, I11. (1983), which describes rezinovye inhibitors containing, methylamino-[CH2NH], and hydroxyethylene-[SNONCE2] bioisostere the amide bond Lei Per 6-13 the oktapeptid derived from angiotensinogen).

In General, the design and synthesis of peptidomimetic that reminds obmenival specificity of ligand/receptor, begins with a sequence obmenivalas specificity of ligand/receptor and data structure (for example, data about geometry, such as bond length and angles between them) desirable obmenivalas specificity of ligand/prescriptions the ora (e.g., most likely the simulated peptide), and use such data to identify geometries that should be embodied in peptidomimetic. To perform this step in the prior art numerous methods and techniques, any of which may be used. (See, for example. Farmer, P.S.Drug Design, (Ariens, E.J. ed.), Vol.10, pp.119-143 (Academic Press, New York, London, Toronto, Sydney and San Francisco) (1980); Farmer, et al., in TIPS, 9/82, pp.362-365; Verber et al., in TINS, 9/85, pp.392-396; Kaltenbronn et al., bj. Med Chem. 33: 838-845 (1990); and A.F. Spatola Chemistry and Biochemistry of Amino Acids. Peptides and Proteins, Vol.7, pp.267-357, Chapter 5, "Peptide Backbone Modifications: A Structure-Activity Analysis of Peptide Containing Amide Bond Surrogates. Conformational Constraints and Relations" (Weistein, B., ed.; Marcel Dekker, New York, pub.) (1983); Kemp, D.S., "Peptidomimetics and the Template Approach to Nucleation of 3-sheets and ∀-helices in Peptides", Tibech, Vol.8, pp.249-255 (1990)). Additional materials can be found in U.S. patents№5288707; 5552534; 5811515; 5817626; 5817879; 5821231 and 5874529. When peptidomimetic constructed, it can be obtained using conventional in peptide chemistry and/or organic chemistry technologies.

Some embodiments of the contain multiple domains specificity and/or more antigenic domains. One of the types of obmenivatsa specificity of ligand/receptor having multiple domains specificity and/or antigenic domains, called "multipersoona obmanivali specificity of ligand/receptor", as they have several domain specificity or antigenic domains, which appear in tandem on the same molecule. For example, multimediabay domain specificity may contain two or more ligands that interact with a single type of receptor, two or more ligands that interact with different types of receptors on the pathogen, and two or more ligands that interact with different types of receptors on different pathogens.

Similarly, multimediabay antigenic domain can be designed to contain multimeric the same epitope of a pathogen or different epitopes of the pathogen, which can also be multimerization. That is, some multipersoona antigenic domains are multivalent, since the same epitope repeats. On the contrary, some multipersoona antigenic domains have more than one epitope that is present on the same molecule in tandem, but different epitopes. Data antigenic domains are multipersonality, but not polyvalent. Additionally, some multipersoona antigenic domains are constructed so as to have different epitopes, but these different epitopes are multivalent, because each type epitope is repeated.

Some obmanivali specificity of ligand/receptor contain other elements in addition to the postponing the specificity and antigenic domain, such as sequences that facilitate purification, linkers, providing more flexibility and reducing the volumetric obstacles, and sequences that either provide more stability obmenivalas specificity of ligand/receptor (e.g., resistance proteasome destruction), or contribute to the destruction (for example, the recognition sites of the protease). For example, obmanivali specificity of ligand/receptor may contain cleaved signal sequences that promote the cytoplasmic export of the peptide, and/or split label sequence that facilitates purification or antibody-based test in columns, glutathione columns and metal columns.

Obmanivali specificity of ligand/receptor may contain elements that contribute to the flexibility of the molecule, reducing volumetric obstacles or allow obmenival specificity of ligand/receptor attached to the base or another molecule. These elements together are called "linkers". One of the types of linkers that can be introduced in obmenival specificity of ligand/receptor is, for example, avidin or streptavidin (or their ligand - Biotin). Through the Biotin-avidin/streptavidin connection several obmenivatsa specificity of ligand/receptor can be connected to each other (for example,through the base, such as molasses, or directly), or you can connect a separate domain specificity and antigenic domains. Another example of a linker, which may be included in obmenival specificity of ligand/receptor, called the "8-linker", as it has a sequence that is found in the phage 8. Preferred 8 sequences are those that correspond to the flexible branches of the phage. These sequences can be entered in obmenival specificity of ligand/receptor (e.g., between domain specificity and antigenic domain or between multimarine domain specificity and/or antigenic domains) to provide greater flexibility and reduce the volume of the obstacle.

Additionally, obmanivali specificity of ligand/receptor may contain sequences that either provide resistance proteasome destruction, or contribute proteasome destruction. By introducing several cysteines in obmenival specificity of ligand/receptor may be achieved, for example, a large resistance proteasome destruction. It is expected that these embodiments of obmenivalas specificity of ligand/receptor remain in the body for long periods, which may be advantageous for some therapeutic applications. On the contrary, obmanivali JV is civicnet ligand/receptor may also contain sequences which contribute to the rapid destruction, to promote rapid removal from the body. Many sequences that serve as recognition sites for serine, cysteine and aspartic proteases, are known and can be entered in obmenival specificity of ligand/receptor.

The following section describes in more detail the domain specificity.

Domains specificity

Types of domain specificity, which can be used in obmanivali specificity of ligand/receptor, diverse, because there are a large number of ligands that interact with receptors on the bacteria, parasites, fungus, mold, viruses and cancer cells. Many types of bacteria, parasites, fungus, mold, viruses and cancer cells, for example, interact with extracellular matrix proteins. Thus, the desired domain specificity contain at least one ligand which has a peptide sequence present in the extracellular matrix protein. That is, the domain specificity may contain a ligand, a peptide having the sequence to be detected, for example, fibrinogen, collagen, vitronectin, laminin, plasminogen, thrombospondin and fibronectin.

Researchers have identified areas of extracellular matrix proteins that interact with multiple R is the receptors. (See, for example, McDev-vit et al., Eur. J. Biochem., 247:416-424 (1997); Flock, Molecular Med. Today, 5:532 (1999); and Pei et al., Infect and Immun. 67:4525 (1999)). Some receptors are associated with the same plot extracellular matrix protein, some have overlapping binding domains, and some are linked together in different areas. Preferably, the ligands comprising the domain specificity, have an amino acid sequence that has been identified as involved in adhesion to extracellular matrix protein. However, it should be understood that random fragments of known ligands for any of the receptor on the pathogen can be used to establish obmenivatsa specificity of ligand/receptor, and these obmanivali candidates specificity of ligand/receptor can be examined during characteristic tests, described below, to identify molecules that interact with receptors on the pathogen.

Some domains specificity contain a ligand that interacts with bacterial adhesion receptor, including, in no particular order restrictions, extracellular fibrinogen binding protein (Efb), linking the collagen protein, vitronectin binding protein, laminin binding protein, plasminogen binding protein, binding thrombospondin protein aggregation factor A (ClfA), an aggregating factor B (ClfB), St. the fibronectin binding protein, the coagulase and extracellular protein adhesion. It is shown that the ligand having the amino acid sequence corresponding to the C-terminal part of the gamma chain of fibrinogen, competitive inhibit the binding of fibrinogen with ClfA, receptor adhesion of Staphylococcus aureus. (McDevvit et al., Eur. J. Biochem., 247:416-424 (1997)). Further, the organisms Staphylococcus produce much more of these adhesion receptors, as Efb, which are associated with fibrinogen alpha chain, ClfB, which interacts with αand β chains of fibrinogen, and Fbe, which is associated with β chain of fibrinogen. (Pei et al., Infect. And lmmun. 67:4525 (1999)). Accordingly, the preferred domain specificity contain at least 3 amino acids from the sequence present in the molecule (e.g., fibrinogen), which can communicate with bacterial adhesion receptor.

The domain specificity may also contain a ligand that interacts with the viral receptor. There are several viral receptors and their respective ligands, these ligands or fragments thereof can be embedded in obmenival specificity of ligand/receptor. For example, Tong et al., identified receptor hepadnavirus, 170-kd-cell surface glycoprotein that interacts with the domain pre-S protein shell of the virus duck hepatitis B (U.S. Patent No. 5929220), a Maddon et al., has determined that the surface protein CD4 T-cells (or soluble form, called T4) interacts with gp120 of HIV (U.S. Patent No. 6093539). Thus, the domain specificity of interacting with the viral receptor sites domain pre-S protein shell of the virus duck hepatitis B (for example, amino acid residues or 80-102 80-104) or sections of the surface protein CD4 T-cells (or soluble form, called T4), interacting with gp120 of HIV (e.g., the extracellular domain of CD4/T4 or its fragments). There are many viral ligands for receptors, and these molecules or their fragments can be used as domain specificity.

The domain specificity may also contain a ligand that interacts with a receptor present on a cancer cell. Protooncogene HER-2/neu (C-erbB2) encodes factor receptor surface growth of a family of tyrosine kinases, p185HER2. Twenty to thirty percent of patients with breast cancer purexpression the gene encoding HER-2/neu (C-erbB2), by gene amplification. Thus, obmanivali specificity of ligand/receptor, containing the domain specificity, which encodes the ligand for HER-2/neu (C-erbB2), are desirable options perform. Many types of cancer cells also purexpression or differentially Express integranova receptors. Many preferred embodiments of the contain the domain specificity, which vzaimode is istue with integranova receptor. Although integrins to the greatest extent interact with extracellular matrix proteins, it is known that these receptors interact with other ligands, such as invasin, peptides containing the RGD (arginine-glycine-aspartate) and chemicals. (See, for example, U.S. patent No. 6090944 and 6090388; Brett et al., Eur J Immunol, 23:1608 (1993)). Ligands for integranova receptors contain, not in order restrictions, molecules reacting with vitronectin receptor, leninova receptor, fibronectin receptor, collagen receptor, fibrinogenesis receptor, receptor α4β1, receptor α6β1, receptor α3β1, receptor α5β1and receptor αvβ3. Table 1 also shows several preferred domain specificity. The above domain specificity are provided for illustrative purposes only, and in no way should be construed as limiting the scope of domain specificity, which can be used in embodiments described herein execution.

The following section describes in more detail antigenic domains.

Table 1

DOMAINS SPECIFICITY
YGEGQQHHLGGAKQAGDV(SEQ.ID.No.1)
MSWSLHPRNLILYFYALLFL (SEQ.ID.No.2)
ILYFYALLFLSTCVAYVAT(SEQ.ID.No.3)
SSTCVAYVATRDNCCILDER(SEQ.ID.No.4)
RDNCCILDERFGSYCPTTCG(SEQ.ID.No.5)
FGSYCPTTCGIADFLSTYQT(SEQ.ID.No.6)
LADFLSTYQTKVDKDLQSLE(SEQ.ID.No.7)
KVDKDLQSLEDILHQVENKT(SEQ.ID.No.8)
DILHQVENKTSEVKQLIKAI(SEQ.ID.No.9)
SEVKQLIKAIQLTYNPDESS(SEQ.ID.No.10)
QLTYNPDESSKPNMIDAATL(SEQ.ID.No.11)
KPNMIDAATLKSRIMLEEIM(SEQ.ID.No.12)
KSRIMLEEIMKYEASILTHD(SEQ.ID.No.13)
KYEASILTHDSSIRYLQEIY(SEQ.ID.No.14)
SSIRYLQEIYNSNNQKIVNL(SEQ.ID.No.15)
NSNNQKIVNLKEKVAQLEAQ(SEQ.ID.No.16)
CQEPCKDTVQIHDITGKDCQ(SEQ.ID.No.17)
IHDITGKDCQDIANKGAKQS(SEQ.ID.No.18)
DIANKGAKQSGLYFIKPLKA(SEQ.ID.No.19)
GLYFIKPLKANQQFLVYCEI(SEQ.ID.No.20)
NQQFLVYCEIDGSGNGWTVF(SEQ.ID.No.21)
DGSGNGWTVFQKRLDGSVDF(SEQ.ID.No.22)
QKRLDGSVDFKKNWIQYKEG(SEQ.ID.No.23)
KKNWIQYKEGFGHLSPTGTT(SEQ.ID.No.24)
FGHLSPTGTTEFWLGNEKIH(SEQ.ID.No.25)
EFWLGNEKIHLISTQSAIPY(SEQ.ID.No.26)
LISTQSAIPYALRVELEDWN(SEQ.ID.No.27)
ALRVELE DWNGRTSTAYAM (SEQ.ID.No.28)
GRTSTADYAMFKVGPEADKY(SEQ.ID.No.29)
FKVGPEADKYRLTYAYFAGG(SEQ.ID.No.30)
RLTYAYFAGGDAGDAFDGFD(SEQ.ID.No.31)
DAGDAFDGFDFGDDPSDKFF(SEQ.ID.No.32)
FGDDPSDKFFTSHNGMQFST(SEQ.ID.No.33)
TSHNGMQFSTWDNDNDKFEG(SEQ.ID.No.34)
WDNDNDKFEGNCAEQDGSGW(SEQ.ID.No.35)
NCAEQDGSGWWMNKCHAGHL(SEQ.ID.No.36)
WMNKCHAGHLNGVYYQGGTY(SEQ.ID.No.37)
NGVYYQGGTYSKASTPNGYD(SEQ.ID.No.38)
SKASTPNGYDNGIIWATWKT(SEQ.ID.No.39)
NGIIWATWKTRWYSMKKTTM(SEQ.ID.No.40)
RWYSMKKTTMKIIPFNRLTI(SEQ.ID.No.41)
KIIPFNRLTIGEGQQHHLGGAKQAGDV(SEQ.ID.No.42)

Antigenic domains

The diversity of antigenic domains, which can be used in obmenivatsa specificity of ligand/receptor, also very great, because the pathogen or toxin may have a variety of imitators. That is antigenic domains, which can be embedded in obmenival specificity of ligand/receptor include the epitopes present in bacteria, fungi, plants, mold, viruses, cancer cells and toxins. Desirable antigenic domains contain the epitope of a pathogen that is already present in the subject due to purchase the Tenno naturally immunity or vaccination. Epitopes of pathogens, for example, childhood diseases, can be used as antigenic domains.

Some perform contain antigenic domain that interacts with the antibody, which was introduced to the subject. For example, the antibody that interacts with antigenic domain on obmanivali specificity, may be injected together with obmenivalas specificity. Further, the antibody that interacts with obmenivalas specificity of ligand/receptor, in a usual condition does not exist in the subject, but the subject gets this antibody by introducing a biological material (for example, serum, blood or tissue). For example, subjects who made blood transfusion, acquire numerous antibodies, some of which can interact with the antigenic domain of obmenivalas specificity of ligand/receptor. Some preferred antigenic domains for use in obmanivali specificity of ligand/receptor contain viral epitopes, including, in no particular order restrictions, herpes simplex virus, hepatitis B virus, TT virus and polio.

In some embodiments, execution also preferably, the antigenic domains contain an epitope of a pathogen or toxin, which recognized "the antibody with high titer". Approaches to the definition of race is uznavaemosti epitope of a pathogen or toxin antibody with high titer described below. To epitopes of a pathogen that can be included in the antigenic domain of obmenivalas specificity of ligand/receptor include epitopes on the peptide sequences disclosed in the patent Sweden No. 9901601-6; U.S. patent No. 5869232; Mol. Immunol. 28:719-726 (1991); and J. Med. Virol. 33:248-252 (1991). Table II shows the amino acid sequences of several preferred antigenic domains.

The section after the Table II describes the preparation of obmenivatsa specificity of ligand/receptor in more detail.

Table II

ANTIGENIC DOMAINS
GLYSSIWLSPGRSYFET (SEQ.ID.No.43)
YTDIKYNPFTDRGE6NM (SEQ.ID.No.44)
DQNIHMNARLLIRSPFT (SEQ.ID.No.45)
LIRSPFTDPQLLVHTDP (SEQ.ID.No.46)
QKESLLFPPVKLLRRVP (SEQ.ID.No.47)
PALTAVETGAT (SEQ.ID.No.48)
STLVPETT (SEQ.ID.No.49)
TPPAYRPPNAPIL (SEQ.ID.No.50)
EIPALTAVE (SEQ.ID.No.51)
LEDPASRDLV (SEQ.ID.No.52)
HRGGPEEF (SEQ.ID.No.53)
HRGGPEE (SEQ.ID.No.54)
VLICGENTVSRNYATHS (SEQ.ID.No.55)
KINTMPPFLDTELTAPS (SEQ.ID.No.56)
PDEKSQREILLNKIASY SEQ.ID.No.57)
TATTTTYAYPGTNRPPV (SEQ.ID.No.58)
STPLPETT (SEQ.ID.No.59

Ways of making obmenivatsa specificity of ligand/receptor that interacts with receptors on the bacteria, parasites, fungi, mold, viruses, and cancer cells

In some embodiments, performing the domain specificity and antigenic domains are manufactured separately and then joined to each other (e.g., via linkers, or by binding to a common carrier molecule), while in other versions of the domain specificity and antigenic domain are made as parts of the same molecule. For example, any of the domain specificity, listed in Table I, can connect with any of the antigenic domains in Table II. Although the domain specificity and antigenic domains can be manufactured separately and connected to each other through a linker or a carrier molecule (e.g., a complex containing biotinylated domain specificity, streptavidin and biotinylated antigenic domain), it is preferable that obmenival specificity of ligand/receptor has been made as a fusion protein. Thus, the preferred options for implementation include fusion proteins containing any of the domain specificity, listed in Table I, the United States with any of the antigenic domains from Table II.

Obmanivali specificity of ligand/receptor can be produced in accordance with conventional methods of protein engineering, protein chemistry, organic chemistry and molecular biology. Additionally, some businesses produce custom-made peptides, and obmenival specificity of ligand/receptor may be obtained by providing such a company sequence desired obmenivalas specificity of ligand/receptor and the use of their services for the production of an agent in accordance with the specifications (e.g., Bachem AG, Switzerland). Preferably, obmanivali specificity of ligand/receptor are prepared by using methods of chemical synthesis such as solid phase peptide synthesis) using known from the prior art methods, such as described in Merrifield et al., J. Am. Chem. Soc. 85:2149 (1964), Houghten et al., Proc. Natl. Acad. Sci. USA, 85:51:32 (1985), Stewart and Young (Solid Phase peptide synthesis. Pierce Chem Co., Rock-ford, IL (1984) and Creighton, 1983, Proteins: Structures and Molecular Principles, W.H. Freeman & Co., N.Y.

In accordance with one approach of solid-phase peptide synthesis is performed using a peptide synthesizer, Applied Biosystems 430A (Applied Biosystems, Foster City, CA). Each synthesis uses R-methylbenzhydrylamine solid-phase carrier resin (Peptide International, Louisville, KY), outstanding carboxyl-terminal amide in the removal of pepti the s from a solid Foundation using acid hydrolysis. Before using carboxyl-terminal amide can be removed, and obmanivali specificity of ligand/receptor can be purified by high performance liquid chromatography (for example, obetovannoi high-performance liquid chromatography (RP-WPGH) using columns PepS-15 18 (Pharmacia, Uppsala, Sweden)), and the sequence is determined by the determinant peptide sequence Applied Biosystems 473A. An alternative synthetic approach uses an automated peptide synthesizer (Syro, Multisyntech, tübingen, Germany) and protected 9-toradoltoradol (Fmoc) amino acids (Milligen, Bedford, MA).

Although obmanivali specificity of ligand/receptor can be synthesized chemically, may be more effective to get these polypeptides by recombinant DNA technology using well-known in the prior art methods. Such methods can be used to construct expression vectors containing nucleotide sequences encoding obmenival specificity of ligand/receptor and appropriate transcriptional and translational control signals. These methods include, for example, methods in vitro recombinant DNA and synthetic techniques and genetic recombination in vivo. Alternatively, RNA capable of encoding obmenival specificity Li gang is a/receptor, can be synthesized chemically using, for example, synthesizers. See, for example, the methods described in Oligonucleotide Synthesis, 1984, Gait, M.J. ed., IRL Press, Oxford.

Many vector systems expresii in the host can be used for the expression of obmenivatsa specificity of ligand/receptor. If obmenival specificity of ligand/receptor is a soluble molecule, it can be recovered from the culture, that is, from the host cell in cases where the peptide or polypeptide is not secretiruetsa, and cultural environment in cases where the peptide or polypeptide secretiruetsa cells. However, the expression systems also encompass engineered host cell that Express associated with membrane obmanivali specificity of ligand/receptor. Purification or enrichment of obmenivatsa specificity of ligand/receptor from such expression systems can be performed using appropriate detergents and lipid micelles and methods well known to specialists.

To expression systems that can be used include, in no particular order constraints, such microorganisms as bacteria (for example E. coli or .subtilis), transformed using expression vectors recombinant DNA bacteriophage, plasmid DNA or kosmidou DNA containing the nucleotide sequence encoding the transfer of ivatel specificity of ligand/receptor; yeast (for example Saccharomyces, Pichia}, transformed with recombinant yeast expression vectors containing the nucleotide sequence encoding obmanivali specificity of ligand/receptor; cell system of insects infected with recombinant virus expression vectors (e.g. Baculovirus)containing the nucleic acid encoding obmanivali specificity of ligand/receptor; or cellular system mammalian (e.g., COS, Cho, KSS, 293, 3T3)containing a recombinant expression constructs containing nucleic acids encoding obmanivali specificity of ligand/receptor.

In bacterial systems a number of expression vectors may be advantageous to selected depending on the intended for obmenivalas specificity of ligand/receptor use. For example, when it is desirable large number (for example, for the production of the pharmaceutical composition of obmenivatsa specificity of ligand/receptor)can be desirable, vectors that direct the expression of high levels of fusion protein products that are already refined. Such vectors include, not in order of restriction, the expression vector pUR278 E.coli (Ruther et al., EMBO J. 2:1791 (1983), in which the coding sequence of obmenivalas specificity of ligand/re is aptara can be individually lagerbuchse into the vector in frame with coded plot lacZ so, to get a fusion protein; pIN vectors (Inouye &Inouye, Nucleic Acids Res., 13:3101-3109 (1985); Van Heeke &Schuster, J. Biol. Chem., 264:5503-5509 (1989)); and the like. Vectors of SUCH can also be used to ekspressirovali foreign polypeptides as fusion proteins with glutathione S-transferase (GST). In General, such fusion proteins are soluble and can be purified from lysed cells by adsorption to glutathione-agarose beads followed by elution in the presence of free glutathione. Vectors of SUCH designed to contain protease sites of cleavage by thrombin or factor XA, so that the cloned target gene product can be released from the share of GST.

In nasekomoe system uses the virus nuclear polyhedrosis Autographa californica (AcNPV) as a vector for the expression of alien genes. The virus grows in the cells of Spodoptera frugiperda. Gene the coding sequence of obmenivalas specificity of ligand/receptor can be cloned separately in the optional areas (for example, in gene polyhedrin) virus and falls under the control of the AcNPV promoter (for example, polikandriotis promoter). Successful insertion of the gene coding sequence obmenivalas specificity of ligand/receptor will lead to deactivation of the gene polyhedrin and obtaining unclosed recombinant virus (i.e., the virus that no is there protein floor, the encoded gene polyhedrin). These recombinant viruses are then used to infect cells of Spodoptera frugiperda, in which is expressed the inserted gene. (For example, see Smith et al., J. Virol. 46:584 (1983); and Smith, U.S. patent No. 4215051).

In mammalian cells the host can be set based on virus expression systems. In those cases, when the expression vector is adenovirus, the sequence of the nucleic acid encoding obmenival specificity of ligand/receptor, can be Legerova with adenovirus transcription/translation control complex, for example, the late promoter and tripartite leading sequence. This chimerical gene can then be inserted in the adenovirus genome by recombination in vitro or in vivo. Inserting in the optional section of the viral genome (e.g., plot E1 or E3) will result in a recombinant virus that is viable and able to Express the gene product of obmenivalas specificity of ligand/receptor in infected hosts. (See, for example, Logan &Shenk, Proc. Natl. Acad. Sci. USA 81:3655-3659 (1984)). Specific trigger signals may also be required for efficient translation of inserted nucleotide sequences obmenivalas specificity of ligand/receptor (e.g., iniziare is the overall codon ATG and related sequences). In most cases, should be provided with exogenous translational control signals, including the initiating codon ATG. Moreover, the initiating codon must be in phase with the reading frame desired coding sequence to ensure translation of the entire insert. These exogenous translational control signals and initiating codons can be of different nature, both natural and synthetic. The efficiency of expression can also be enhanced through the implementation of appropriate enhance transcription elements, breakers transcription and so on (See. Bittner et al., Methods in Enzymol., 153:516-544 (1987)).

In addition, you may be selected strain of host cells that modulates the expression of the inserted sequences, or modifies and processes the gene product is desired in a specific way. Such modifications (e.g., glycosylation) and processing (e.g., cleavage) of protein products may be important in some embodiments, execution. Different host cells have characteristic and specific mechanisms for pokerenlinea processing and modification of proteins and gene products. Appropriate cell lines or system owner can be chosen to ensure the correct modification and processing of the expressed alien protein. For this you may use the change eukaryotic host cell, with cellular machinery for proper processing of the primary transcript, glycosylation, and phosphorylation of the gene product. Such mammalian cell hosts include, in no particular order restrictions, Cho, VERO, BHK, HeLa, COS, MDCK, 293, 3T3, and WI38.

For long-term high-performance production of recombinant proteins preferred stable expression. For example, can be developed cell lines that stably Express the above obmanivali specificity of ligand/receptor. Rather than using expression vectors that contain viral sources of replication, host cells can be transformed with DNA controlled by suitable controls expression (e.g., promoter, enhancing sequences, breakers transcription sites of polyadenylation etc), and selective marker. After the introduction of alien DNA, engineered cells to grow for 1-2 days in an enriched medium, and then transferred to selective medium. Selective marker in the recombinant plasmid gives resistance to the selection and allows cells to stably integrate the plasmid into their chromosomes and grow, forming foci, which, in turn, cloned and propagated in cell lines. This method is preferably used is to develop lines, expressing obmenival specificity of ligand/receptor.

You can use a variety of systems, including, in no particular order restrictions, genes timedancing of herpes simplex virus (Wigler, et al., Cell 11:223 (1977)), gipoksantin-guanine phosphoribosyltransferase (Szybalska &Szybalski, Proc. Natl. Acad. Sci. USA 48:2026 (1962)), and adenine-phosphoribosyltransferase (Lowy, et al., Cell, 22:817 (1980)) can be used respectively in the cells of tk. sup-, hgprt. sup - or aprt. sup-. Antimetabolite resistance can also be used as the basis of selection for the following genes: dhfr, which gives resistance to methotrexate (Wigler, et al., Proc. Natl. Acad. Sci. USA 77:3567 (1980)); O'hare et al., Proc. Natl. Acad. Sci. USA 78:1527 (1981)); gpt, which gives resistance mycophenolic acid (Mulligan &Berg, Proc. Natl. Acad. Sci. USA 78:2072 (1981)); neo, which gives resistance to the aminoglycoside G-418 (Colberre-Garapin, et al., J. Mol. Biol. 150:1 (1981)); and hygro, which gives resistance to hygromycin (Santerre, et al., Gene 30:147 (1984)).

The following section describes the characteristic tests of obmenivalas specificity of ligand/receptor in more detail.

Characteristic tests obmenivalas specificity of ligand/receptor

Preferably obmanivali specificity of ligand/receptor analyzed for the ability to interact with the receptor and/or the ability to interact with the antibody, which can prists who participate in the subject. The term "characteristic test" means an analysis, experiment or test performed on obmenivalas specificity of ligand/receptor, which evaluates the ability of obmenivalas specificity of ligand/receptor to interact with the receptor (for example, a surface receptor that is present in bacteria, viruses, mold, or fungi) or antibody (e.g., an antibody that recognizes an epitope on the pathogen), or to have an impact on the proliferation of the pathogen. The term "characteristic test" covered research associate (e.g., enzyme immunoassy (fit), enzyme-linked immune tests (ELISA)tests of competitive binding, computer-designed test link test link with a basis of one and dvuhkabinnye system) and studies of infectivity (e.g., reduce viral infection, propagation, and attachment to the host cell).

Preferred tests use binding multimeric agents. One of the forms a multimeric agent associated with the composition containing obmenival specificity of ligand/receptor or its fragments placed on the canvas. Another form multimeric agent comprises a composition containing a receptor or antibody specific for the antigenic domain of obmenivalas specificity of ligand/receptor located on the fundamentals of the. "Basis" can be a carrier, protein, resin, cell membrane, or any macromolecular structure used to connect or immobilization of these molecules. The solid bases include, in no particular order restrictions, the walls of the cells of the reaction tray, test tubes, polystyrene beads, magnetic beads, nitrocellulose strips, membranes, such microparticles, such as latex particles, animal cells, Duracyte®, artificial cells, and so on. Obmenival specificity of ligand/receptor may also be connected with such inorganic bases as silicon oxide material (e.g. silica gel, zeolite, diatomaceous earth or aminirovanie glass), for example through covalent binding through a hydroxy, carboxy or amino group and the reactive group-based.

In some multimeric agents macromolecular base has a hydrophobic surface that interacts with a part of obmenivalas specificity of ligand/receptor, receptor or ligand by hydrophobic non-covalent interaction. In some cases, the hydrophobic surface of the base is such a polymer, such as plastic, or any other polymer, in which the hydrophobic groups are linked, such as polystyrene, polyethylene or polyvinyl. Additionally, obmenival specificity League is Yes/receptor, the receptor or antibody specific for the antigenic domain of obmenivalas specificity of ligand/receptor may be covalently linked with the basics, including proteins and oligo/polysaccharides (e.g. cellulose, starch, glycogen, chitosane or zaminirovannoy separate). In these last multimeric agents reactive group on the molecule, such as hydroxy - or amino group, is used for compounds with a reactive group on the carrier to create the covalent bond. Additional multimeric agents contain the basis having other reactive groups that are chemically activated so as to attach obmenival specificity of ligand/receptor, receptor or antibody, specific for the antigenic domain of obmenivalas specificity of ligand/receptor. Can for example be used CYANOGEN-bromide-activated matrix, epoxy-activated matrix, thio - thio-propyl gels, nitrophenylphosphate and N-hydroxy - succinimide-chloroformate ligament or exernally basis. (Sigma). Further, in some embodiments, performing as a basis, it is assumed liposome or lipid bilayer (natural or synthetic), and obmenival specificity of ligand/receptor, receptor or antibody, specific for the antigenic domain of obmenivalas specificity of ligand/receptor, m which can be attached to the membrane surface or incorporated into the membrane using methods liposomal engineering. In accordance with one approach liposomal multimeric bases contain obmenival specificity of ligand/receptor, receptor or antibody, specific for the antigenic domain of obmenivalas specificity of ligand/receptor exposed on the surface.

Also refers to the insertion of linkers (e.g., "λ-linkers", designed so that they resembled the flexible parts of the phage) of an appropriate length between obmenivalas specificity of ligand/receptor, receptor or antibody, specific for the antigenic domain of obmenivalas specificity of ligand/receptor and the framework to provide greater flexibility and to overcome any surround obstacles that involves basis. Determining the appropriate length of linker that allows optimal binding can be accomplished by screening attached molecules with different linkers in detail described here are characteristic tests.

Several approaches to characterization obmenivatsa specificity of ligand/receptor use multimer described above. For example, associated with the basis obmenival specificity of ligand/receptor may come in contact with "free" adhesion receptors, and the relationship can be determined directly (e.g., by using marterbating adhesion receptors) or indirectly, for example, using the labeled ligand directed to the adhesion receptor). Thus, the candidate obmanivali specificity of ligand/receptor identified these obmenivatsa specificity of ligand/receptor by binding receptors linked with the basis of the candidate obmenivalas specificity of ligand/receptor. The alternative associated with the base adhesion receptors may come into contact with "free" obmenivatsya specificity of ligand/receptor, and the number of associated obmenivalas specificity of ligand/receptor can be determined directly (e.g., using bulleted obmenivalas specificity of ligand/receptor) or indirectly (for example, using a labeled antibody directed to the antigenic domain of obmenivalas specificity of ligand/receptor). Similarly, using antibodies specific for the antigenic domain of obmenivalas specificity of ligand/receptor-based, and bulleted obmenivalas specificity of ligand/receptor (or secondary detectiong reagent, for example, the labeled receptor or antibodies to obmenival specificity of ligand/receptor), may be determined by the ability of antibodies to contact antigenic domain obmenivalas specificity of ligand/receptor.

Some characteristic tests evaluate the ability of obmenivalas specificity of ligand/receptor to interact with a target receptor and redirects antibody, while other characteristic tests designed to determine whether obmenival specificity of ligand/receptor to communicate with the target receptor, and redirects antibody. In General, the characteristic tests can be classified as (1) characteristic tests in vitro, (2) cell characteristic test, and (3) the characteristic in vivo tests.

The discussion of each type of characteristic tests described in the following sections.

Characteristic tests in vitro

There are many types of in vitro tests that can be used to determine whether the associated obmenival specificity of ligand/receptor with a specific receptor, and whether the antibody in the subject, contact obmenivalas specificity of ligand/receptor. In the simplest embodiment, the receptor is associated with the basis (e.g., Petri dish), and this binding obmenivalas specificity of ligand/receptor with the receptor is monitored directly or indirectly as described above. Similarly, a primary antibody directed to the antigenic domain of obmenivalas specificity of ligand/receptor (the example the antibody in the subject), can be connected with the base, and linking obmenivalas specificity of ligand/receptor primary antibody can be determined directly (e.g., using bulleted obmenivalas specificity of ligand/receptor) or indirectly (for example, using the labeled receptor or the labeled secondary antibodies which are reactive with the epitope on obmanivali specificity of ligand/receptor, which does not compete with the epitope recognized by the primary antibody).

Another approach contains a quiz on the sandwich principle, when the receptor binds to the base, obmenival specificity of ligand/receptor binds to the receptor, and the primary antibody binds to obmenivalas specificity of ligand/receptor. If used with an appropriate primary antibody, the presence of a complex of receptor/obmenival specificity/primary antibody can be determined directly. The presence of a complex of receptor/obmenival specificity/primary antibody can also be determined indirectly using, for example, the labeled secondary antibody, which reacts with the primary antibody for the epitope that does not interfere with the binding of the primary antibodies with obmenivalas specificity of ligand/receptor. In some cases it may be desirable to use m Kirovskoe tertiary antibody, reacting with unlabeled secondary antibody with the formation of so complex receptor/obmenival specificity/primary antibody/secondary antibody/labeled tertiary antibody.

The following example describes the characteristic test was performed to determine whether the domain inhibits specificity, derived from the C-terminal domain of fibrinogen, link aggregation factor (Clf) with fibrinogen.

EXAMPLE 1

In this example, several peptides corresponding to the C-terminal domain of fibrinogen (Fib)were analyzed with respect to their ability to block the binding of an aggregating factor (Clf) with fibrinogen. (See Table III). These peptides were produced using standard peptide synthesis methods using fmoc-chemistry (Syro, MultiSynTech, Germany). Preferably, the peptides are purified by bratovanova WPGH. Then produced a competitive enzyme immune tests to determine whether the peptides capable of blocking the interaction between Clf and fibrinogen. The results of these experiments are shown in Table III. The smallest peptide from fibrinogen, which was able to inhibit the interaction between Clf and fibrinogen was HLGGAKQAGDV (ID. PEFC. No. 124).

Tab the Itza III
SEQ ID NO. (Fib) peptideInhibition of the interaction (Fib/Clf)
106LTIGEGQQHHLGGAKQAGDV+
107GEGQQHHLGGAKQAGDV+
108QQHHLGGAKQAGDV+
109QHHbGGAKQAGDV+
110HHLGGAKQAGDV+
111HLGGAKQAGDV+
112LGGAKQAGDV-
113GGAKQAGDV-
114GAKQAGDV-
115QHHLGGAKQAGD+
116QHHLGGAKQAG+
117QHHLGGAKQA-
118QHHLGGAKQ-
119QHHLGGAK+/-
120QHHLGGA-
121HHLGGAKQAGDV+
122HHLGGAKQAGD+
123HHLGGAKQAG+
124HLGGAKQAGDV+
125HLGGAKQAGD4-
126ALGGAKQAG-
127HAGGAKQAG +
128HLAGAKQAG+
129HLGAAKQAG+
130HLGGGKQAG+
131HLGGAAQAG+/-
132HLGGAKAAG+
133HLGGAKQGG+
134HLGGAKQAA+

The following example describes the characteristic test was performed to determine interacts whether obmenival specificity of ligand/receptor with bacteria with the receptor ClfA.

EXAMPLE 2

Obmanivali specificity of ligand/receptor having the domain specificity (length of approximately 20 amino acids), corresponding to different regions of the sequence of the gamma chain of fibrinogen, were obtained using standard peptide synthesis methods using fmoc-chemistry (Syro, MultiSynTech, Germany), and data obmanivali specificity of ligand/receptor was analyzed in terms of their ability to bind the receptor ClfA and the antibody specific for the antigenic domain. The sequence of these obmenivatsa specificity of ligand/receptor are listed in Table IV, as well as listing sequence ID. PEFC. No. 60-103). Obmanivali specificity of ligand/receptor, usage is implemented in this test, have antigenic domain that represents the epitope gG2 protein of herpes simplex virus, which is recognized by a monoclonal antibody to gG2 protein of herpes simplex virus. Performed serial dilutions of these obmenivatsa specificity of ligand/receptor in phosphate buffer saline (PBS)containing 2:g/ml of goat serum. (Sigma Chemicals, St. Louis, MO) and 0.5% Tween 20 (PBS-GT). Receptor ClfA passively adsorbiroval at 10 µg/ml in 96-cell plate microtitre 50-mmol sodium carbonate buffer, pH of 9.6, overnight at +4°C.

Diluted obmanivali specificity of ligand/receptor was then incubated on the plates for 60 minutes. The ability of obmenivalas specificity of ligand/receptor to interact with the receptor was determined by adding on the plate of the primary antibodies and incubation for 60 minutes (mAb dilution of 1:1000 for gG2 protein of herpes simplex virus). Associated primary mAb was then identified using rabbit antimisting IgG (Sigma) secondary antibodies and peroxidase labeled goat anti-rabbit IgG (Sigma) tertiary antibodies. Plates were revealed by incubation with dinitrobenzenamine (Sigma), and analyzed the absorption at 405 nm.

All obmanivali specificity of ligand/receptor, are shown in Table IV (ID. PEFC. No. 60-103), markedly tie the Wali immobilized ClfA, and also provided the binding of mAb specific for gG2 protein of herpes simplex virus. The above-described method of determining the affinity of obmenivalas specificity of ligand/receptor for adhesion and receptor primary antibodies can be made for any candidate obmenivalas specificity of ligand/receptor containing any domain specificity and any antigenic domain, provided that we will use the appropriate sequence and adhesion receptors.

The example of Table IV describes another characteristic test was performed to determine interacts whether obmenival specificity of ligand/receptor with bacteria with the receptor ClfA.

EXAMPLE 3

Obmanivali specificity of ligand/receptor domains with specificity that are associated with an aggregating factor (Clf), and antigenic domains, which correspond to the epitope derived from polio virus, were obtained using standard peptide synthesis methods using fmoc-chemistry (Syro, MultiSynTech, Germany). Cm. Table V. These obmanivali specificity of ligand/receptor was analyzed in terms of their ability to inhibit the interaction between CLF and fibrinogen. These experiments were carried out describe the data in Table V obmanivali specificity of ligand/receptor, and different concentrations of these molecules were added in the competitive enzyme immunoassay containing Clf and fibrinogen. Installed the lowest inhibitory concentration, the lowest concentration of peptide required to inhibit the interaction Clf/Fib. Accordingly, the lower the concentration required for inhibition of the interaction Clf/Fib, the more effective the inhibitor. Additionally it was determined the lowest concentration of solid-phase bound peptide, which is the lowest tested concentration of a peptide recognized by antibodies to the polio virus, immunotest. Some of the peptides used (for example, CPALTAVETGCTNPLAAHHLGGAKQAG (ID. PEFC. NO. 135), HHLGGAKQAGAACPALTAVETGCTNPL (ID. PEFC. NO. 137), CPALTAVETGCTNPLHHLGGAKQAG (ID. PEFC. No. 139) and HHLGGAKQAGCPALTAVETGCTNPL (ID. PEFC. No. 141))indicated in Table V with asterisks were fixated between two artificially introduced cysteine residues. These experiments showed that HHLGGAKQAG-AA-CPALTAVETGCTNPL (ID. PEFC. No. 137) and HHLGGAKQAG-CPALTAVETGCTNPL (ID. PEFC. No. 142) effectively inhibited the interaction of Clf with fibrinogen and kept functional epitopes of the virus of poliomyelitis.

Table V
The lowest inhibitory conc.The lowest epitope on Tvarditsa
ID Peptide sequence(ág/ml)(ág/ml)
135 CPALTAVETGCTNPL-AA-HHLGGAKQAG*>6251,6
136 CPALTAVETGCTNPL-AA-HHLGGAKQAG6251,6
137 HHLGGAKQAG-AA-CPALTAVETGCTNPL*698
138 HHLGGAKQAG-AA-CPALTAVETGCTNPL625>200
139 CPALTAVETGC-TNPLHHLGGAKQAG*6251,6
140 CPALTAVETGC-TNPLHHLGGAKQAG2081,6
141 HHLGGAKQAG-CPALTAVETGCTNPL*208>200
142 HHLGGAKQAG-CPALTAVETGCTNPL231,6
143 PALTAVETGATNPL-HHLGGAKQAG>6251,6
144 HHLGGAKQAG-PALTAVETGATNPL>625>200

The following section describes several cell-based characteristic tests that can be performed to determine, does obmenival specificity of ligand/receptor effects on the proliferation of the pathogen.

Characteristic tests based on pathogen

Another type of characteristic test is used based on the pathogen approach to assessing the ability of obmenivalas specificity of ligand/receptor to interact with the pathogen and antibody directed by ant the gene domain obmenivalas specificity of ligand/receptor. This test also reveals the ability of obmenivalas specificity of ligand/receptor to influence the proliferation of the pathogen, as in the body of the subject of the interaction of obmenivalas specificity of ligand/receptor with the pathogen and antibody directed towards an antigenic domain of obmenivalas specificity of ligand/receptor, accompanied by humoral and cellular reactions that lead pathogen from a subject (e.g., complement fixation, or destruction by macrophages). In General, based on the pathogen characterization tests include the introduction of obmenivatsa specificity of ligand/receptor in cultured pathogens and tracking link obmenivalas specificity of ligand/receptor with a cell or virus. Can be used several types based on the pathogen characteristic tests, and the following example describes some of the preferred characteristic tests in more detail.

EXAMPLE 4

One type is based on the pathogen characteristic test contains binding obmenivalas specificity of ligand/receptor with bacteria located on the base. Accordingly, the producing bacteria Clfa (e.g., Staphylococcus aureus or Escherichia coli) grown in culture or on agar plate in a suitable growth medium (e.g., LB broth, blood broth, yeah the LB or blood agar). The cells are then transferred to a membrane (e.g. nitrocellulose or nylon) or by placing culture at the membrane under vacuum (for example, using datamatching collector device), or by placing the membrane on the colonies for a period sufficient to transfer. To cells that are associated with the membrane, then served a serial dilution of obmenivalas specificity of ligand/receptor (e.g., 500 ng, 1:g 5:g 10:g 25:g and 50:g obmenivalas specificity of ligand/receptor in a total volume of 200: l FBS). In one experiment was used obmanivali specificity of ligand/receptor listed in Table IV or V. Then diluted obmanivali specificity of ligand/receptor were incubated on the membrane for 60 minutes. Then unbound obmanivali specificity of ligand/receptor are removed and the membrane washed with FBS (for example, 3 wash 2 ml FBS). Then served dilution 1:100-1:1000 primary antibodies which are reactive with antigenic domain obmenivalas specificity of ligand/receptor (e.g., mAb for gG2 protein of herpes simplex virus), and binding assays allow to proceed for 60 minutes. The membrane is washed again with PBS (e.g., 3 wash 2 ml PBS) to remove unbound primary antibody. Suitable control agent which include the membrane, bacteria on the membrane without obmenivalas specificity of ligand/receptor and bacteria on the membrane with obmenivalas specificity of ligand/receptor, but without primary antibody.

To determine the number of obmenivalas specificity of ligand/receptor associated with bacteria on the membrane, used secondary antibody (e.g. rabbit antimachine IgG (Sigma)) and the tertiary antibody (e.g., labeled with peroxidase goat anti-rabbit IgG (Sigma)). Of course, the labeled secondary antibody that interacts with the primary antibody, may also be used. As noted above, the secondary antibody is in contact with the membrane for 60 minutes, and unbound secondary antibody is washed from the membrane with PBS (e.g., 3 wash 2 ml FBS). Then the tertiary antibody is in contact with the membrane for 60 minutes, and unbound antibodies are washed away from the membrane with PBS (e.g., 3 wash 2 ml FBS). Related tertiary antibodies can be detected by incubating the membrane with dinitrobenzenamine (Sigma).

Another approach includes the use of immobilized obmenivalas specificity of ligand/receptor. Accordingly, the primary antibody (for example, mAb for gG2 protein of herpes simplex virus) is associated with the Petri dish. When the primary antibody is due to cover the second Cup, you enter different dilution of obmenivalas specificity of ligand/receptor (e.g., obmenivalas specificity of ligand/receptor, described in Table IV or V). Obmenival specificity of ligand/receptor allow it to connect with the primary antibody for 60 minutes, and unbound obmenival specificity of ligand/receptor washed (for example, 3 wash 2 ml FBS). Suitable control agents include Petri dishes without primary antibodies or obmenivalas specificity of ligand/receptor.

Then the Petri dishes is added to a cloudy solution of bacteria (such as E. coli), and allow bacteria to interact with immobilized obmenivalas specificity of ligand/receptor within 60 minutes. Then unbound bacteria are removed by washing with PBS (e.g., 3 wash 2 ml FBS). Then the growth medium (e.g., LB broth) served in a Petri dish, and the culture incubated overnight. Alternative in a Petri dish is added to the LB agar and the culture incubated overnight. The interaction between obmenivalas specificity of ligand/receptor and bacteria can be observed visually (e.g., turbid environment of growth, which can be measured using spectrophotometric analysis, or the appearance of colonies on the agar).

Modifying the above approaches, the specialist can assess the ability of obmenivalas specificity of ligand/receptor maintains dejstvovatj with the virus. For example, it is shown that soluble fragments of glycoprotein T4 interact with the envelope glycoprotein of human immunodeficiency virus (HIV). (See, for example, U.S. patent No. 6093539). Obmanivali specificity of ligand/receptor, containing the domain specificity, containing a fragment of T4 glycoprotein that interacts with the envelope glycoprotein of HIV (for example, amino acids 1-419 sequence of the T4 glycoprotein described in U.S. patent No. 6093539, or parts of it)can be produced through the synthesis of a fusion protein having a domain specificity, coupled with antigenic domain (e.g., antigenic domain from Table II). Although to obtain obmenivalas specificity of ligand/receptor can be used in peptide chemistry, preferably, the expression construct containing a fragment of glycoprotein T4 connected to the antigenic domain, were made and transferrals in the appropriate cell. Strategies for expression and purification described above and in U.S. patent No. 6093539, may also be used.

When obmenival specificity of ligand/receptor designed, executed test binding to the filter. Accordingly, a serial tenfold dilution of the inoculum of HIV are introduced to the membrane (for example, nitrocellulose or nylon) in the dot-blotterover device at a constant HAC is the mind. Then a serial tenfold dilution of obmenivalas specificity of ligand/receptor are introduced to related HIV particles. Obmenival specificity of ligand/receptor is in contact with the particles within 60 minutes before applying vacuum and washing with PBS (e.g., 3 wash 2 ml FBS). When unbound obmenival specificity of ligand/receptor deleted, serial tenfold dilution of the primary antibody, which binds to the antigenic domain are added to the sample, and binding assays can go for 60 minutes. Then is vacuum, and the unbound primary antibodies, washed with PBS (e.g., 3 wash 2 ml FBS). Associated primary antibodies can be detected as described above.

The ability of obmenivalas specificity of ligand/receptor to interact with the virus can also be evaluated in the analysis according to the sandwich principle. Accordingly, a primary antibody that interact with antigenic domain obmenivalas specificity of ligand/receptor, immobilities cells in microtitre, and serial dilutions of obmenivalas specificity of ligand/receptor are added to the primary antibody, to create a set of primary antibody/obmenival specificity of ligand/receptor, as described above. Then add the using serial ten-fold dilution of the supernatant of HIV, and binding assays can go for 60 minutes. Unlinked HIV particles are removed by successive washes in PBS. The associated discovery of HIV particles can be performed using radiomarimanga antibodies to HIV (for example, antibodies obtained from the serum of a person suffering from HIV infection).

Although the above examples describe based on pathogen tests using bacteria and virus, can be made of modifications of these approaches to study the interaction of obmenivatsa specificity of ligand/receptor from mammalian cells. For example, the ability of obmenivalas specificity of ligand/receptor to interact with integranova receptor present on the cancer cell, can be determined as follows. Melanoma cells expressing the receptor ∀v3(for example, melanoma cells M21 person) bind fibrinogen, and this interaction can be blocked by the filing of a peptide containing the RGD (See, for example, Katada et al., J. Biol. Chem. 272:7720 (1997) and Felding-Habermann et al., J. Biol. Chem. 271:5892-5900 (1996)). Similarly, many other types of cancer cells Express integrins that interact with RGD-peptides. In accordance with one approach cancer cells expressing reacting to RGD integrin (e.g., melanoma cells M21 person), to liverwurst to merge. Cells M21 can be grown in DMEM medium containing 10% serum bovine fetus, 20 mmol Hepes and 1 mmol of pyruvate.

Preferably, the cells are stained by hydroidolina (Polysciences, Inc., Warrington, PA) with a final concentration of 20 µg/ml (2×106cells/ml) for 30 minutes at 37°and then twice washed to remove excess dye. Hydration embedded in DNA, giving the red fluorescent labeling of cells, and does not spoil the adhesive function of cells. Staining provides a way of calculating the binding obmenivalas specificity of ligand/receptor cells. That is, the total number painted hydroidolina cells can be compared with the number of cells associated with fluorescence marked by a complex of primary antibody/obmenival specificity of ligand/receptor, to determine the effectiveness of the binding.

Accordingly, the stained cells are incubated with various dilutions of obmenivalas specificity of ligand/receptor containing the RGD sequence (for example, GRGDSPHRGGPEE (ID. PEFC. No. 104) or WSRGDWHRGGPEE (ID. PEFC. No. 105)). After 60 minutes incubation the unbound obmenival specificity of ligand/receptor is removed by several washes in DMEM medium containing 10% serum bovine fetus 20 mmol Hepes and 1 mmol of pyruvate (for example, three washing in 5 ml of medium). Then served dilution 1:100-1:1000 lane the ranks antibodies interacting with antigenic domain obmenivalas specificity of ligand/receptor (e.g., mAb for gG2 protein of herpes simplex virus), and binding assays can go for 60 minutes. Then run a few washes in the medium to remove unbound primary antibody. Suitable control agents include stained cells without obmenivalas specificity of ligand/receptor or stained cells without primary antibody.

After binding of the primary antibody is added goat antimurine PTS-labeled antibody (dilution 1:100) (Sigma), and the binding of Pets within 60 minutes. Again runs several washes to remove unbound secondary antibody. Is the test by flow cytometry with the installation of the filter on 543/590 nm for hydrocidone and 495/525 nm for fluorescein. You can observe significant binding of the primary antibodies with complex obmenival specificity of ligand/receptor/cell, which demonstrates the fact that obmenival specificity of ligand/receptor affects the cell.

The following example describes the characteristic test, which confirms that RGD-containing obmanivali specificity of ligand/receptor effectively contacted with mammalian cells and redirect antibodies to the t cell.

EXAMPLE 5

Peptide RGDSAATPPAYR (ID. PEFC. No. 145) was prepared using standard peptide synthesis methods using fmoc-chemistry (Syro, MultiSynTech, Germany). This peptide has domain specificity which binds to integranova receptors, the spacer (AA) and antigenic domain that has an epitope recognized by a monoclonal antibody 57/8, the epitope is present on the e-antigen of hepatitis B (HbeAg).

Mouse myeloma cells (cells SP2/0) were washed in serum-free medium, and then were incubated with peptide RGDSAATPPAYR (ID. PEFC. No. 145) or a control peptide derived from the domain of NS3 of hepatitis C (VHC) at a concentration of 50 µg/ml. Then cells were washed, and the amount of bound on the surface of the peptide was determined by labeling cells with antibodies 57/8. Associated on the surface of the antibody was observed in the marked FITC conjugate antimisting IgG, diluted 1/500, and the level of surface staining was determined using fluorescent microscopy.

The experiment showed that cells incubated with the control peptide showed no staining. In contrast, cells incubated with peptide RGDSAATPPAYR (ID. PEFC. No. 145), showed significant surface staining, corresponding to the location of surface-expressed integrins. Accordingly, RGD-containing obmanivali specificity the spine of the ligand/receptor effectively bind generating integrin mammalian cells and these molecules can be used to redirect and targeting antibodies on tumor cells.

The following section describes the characteristic tests on animals.

The characteristic in vivo tests

To characteristic tests are also experiments that assess obmanivali specificity of ligand/receptor in vivo. There are many animal models can be used to assess the ability of obmenivalas specificity of ligand/receptor to inhibit pathogenic infection. Mice are preferred because they are easy to contain, and they are prone to bacterial infections, viral infections and cancer. Chimpanzees also preferred because of their close genetic relationship with people.

The approach to evaluating the effectiveness of obmenivalas specificity of ligand/receptor for mice in the following example.

EXAMPLE 6

To test the ability of obmenivalas specificity of ligand/receptor to treat a bacterial infection, can be performed following characteristic analysis. Several females outbreeding mice CF-1 (Charles Rivers Laboratories) aged approximately 8 weeks and weighing 25 grams was vacciniavirus using antigenic domains of the test obmenivatsa specificity of ligand/receptor. Antigenic domains preferably connected is camping with the media and was injected with the adjuvant. For example, antigenic domains can be fused with "lock-downhole" hemocyanin or albumin bovine serum, which acts as the role of media and the role of adjuvant or can be used adjuvant such as Freund adjuvant, aluminum hydroxide or lysolecithin. As soon as a high titer of antibodies to antigenic domains can be confirmed by using, for example, immunodiffusion or fit immunized mouse inoculants intraperitoneally with grown overnight cultures of Staphylococcus aureus NTCC 10649. The inoculum are selected to give approximately 100×SD50or log 6,6 for S.aureus.

Serial dilutions of obmenivatsa specificity of ligand/receptor (e.g., obmenivatsa specificity of ligand/receptor, are shown in Table IV) are formulated in sterile water for injection and administered by subcutaneous (SC) or oral (PO) route through one and five hours after infection. Simultaneously with each attempt to control the infection SD50was confirmed by inequilibrium untreated mice using logarithmic dilutions of the bacterial inoculum. Preferably, the range of the five logarithmic dilutions of a bacterial infection inoculate five groups of ten mice each (ten mice on a logarithmic dilution). Smart is here 100% will be invoked in all groups of untreated mice control infection inoculate 100× SD50. Mouse tracked daily on the subject of mortality within seven days. The average effective dose to protect 50% of mice (ED50) can be calculated from the total mortality by logarithmic probit analysis plotted the survival curve depending on the dose, as described in Antimicrob. Agents Chemother. 31:1768-1774 and Proc. Soc. Exp.Biol. Med. 1994, 57, 261-264. As noted by the expert, such approaches can be used to test the ability of obmenivatsa specificity of ligand/receptor to inhibit viral infection and cancer.

Described here obmanivali specificity of ligand/receptor can be formulated in the form of pharmaceutical preparations and administered to subjects in need of the agent, inhibiting the proliferation of a pathogen. The following section describes several pharmaceutical preparations containing obmanivali specificity of ligand/receptor that interacts with a receptor on the pathogen.

Pharmaceutical preparations containing obmenival specificity of ligand/receptor that interacts with a receptor on the pathogen

Described here obmanivali specificity of ligand/receptor suitable for introduction into pharmaceutical preparations for administration to subjects in need of the compound that treats or prevents infection by the pathogen. These pharmaceutically active connection is tion can be processed in accordance with generally accepted ways galenical pharmacology for medical agents for the introduction of a mammal, including people. The active ingredients can penetrate into the pharmaceutical product with modification and without it. Further, the manufacture of pharmaceuticals or therapeutic agents that deliver pharmacologically active compounds of the present invention by several routes, an aspect of the present invention. For example, and not in the order of limitation, DNA, RNA, and viral vectors having a sequence encoding obmenival specificity of ligand/receptor that interacts with a receptor on the pathogen, used with variants of carrying out the invention. Nucleic acids encoding obmenival specificity of ligand/receptor may be administered alone or in combination with other active ingredients.

Connections may be used in mixture with conventional excipients, i.e. pharmaceutically acceptable organic or inorganic substances-carriers suitable for parenteral, enteral (e.g. oral) or local injection, so as not to react aggravating way described here pharmacologically active ingredients. Suitable pharmaceutically acceptable carriers include, in no particular order restrictions, water, salt solutions, alcohols, gum Arabic, vegetable oils, benzyl alcohols, polietileno the Oli, gelatin, carbohydrates such as lactose, amylose or starch, magnesium stearate, talc, silicic acid, viscous paraffin, cosmetic oil, monoglycerides and diglycerides of fatty acids, pentaerythritol esters of fatty acids, hydroxymethylcellulose, polyvinylpyrrolidone, etc. more potential holders described in Remmington's Pharmaceutical Sciences, 15thEdition, Easton:Mack Publishing Company, pp.1405-1412 and 1461-1487 (1975) and The National Formulary XIV, 14thEdition, Washington, American Pharmaceutical Association (1975). The pharmaceutical preparations can be sterilized and, if desired, mixed with auxiliary agents such as lubricants, preservatives, stabilizers, wetting, emulsifying agents, salts for influencing osmotic pressure, buffers, coloring, flavoring and/or aromatic substances and the like, if auxiliary agents do not react worsen with obmenivatsya specificity of ligand/receptor.

The effective dosage and route of administration of a particular pharmaceutical product containing obmenival specificity of ligand/receptor can vary depending on the individual needs of the patient and the desired therapeutic or prophylactic purposes. Therapeutic efficacy and toxicity of such compounds can be determined by standard pharmaceutical procedures in cell is ultura or experimental animals, for example the ED50(the dose therapeutically effective in 50% of the population). For example, an effective dose of obmenivalas specificity of ligand/receptor may be assessed using the above characteristic tests. The data obtained in these analyses are then used in formulating a range of dosage for use with other organisms, including people. The dosage of such compounds is preferably in the range of circulating concentrations, including the ED50without toxicity. The dosage varies within this range depending on the type of obmenivalas specificity of ligand/receptor used dosage forms, the sensitivity of the organism and route of administration.

Normal dosage of obmenivalas specificity of ligand/receptor can vary from approximately 1 to 100,000 micrograms, up to a total dose of about 10 grams, depending on the route of administration. The desired dose contains approximately 250:g - 1 mg, about 50 mg to 200 mg and about 250 mg to 500 mg

In some embodiments, execution dose obmenivalas specificity of ligand/receptor preferably gives the concentration in the tissue or blood or there is approximately from 0.1:Mol to 500 mmol. The desired dose given concentration in the tissue or blood or there, equal to approx is approximately from 1 to 800 µmol. Preferred doses given concentration in tissue or blood, greater than from about 10 µm to about 500:Mol. Although the dose, giving the concentration in the tissue is greater than 800:Mol, are not preferred, they can be used. Also can be provided by a constant infusion of obmenivalas specificity of ligand/receptor, in order to maintain a stable concentration in the tissues by measuring levels in the blood.

The exact dosage is selected by a specific physician, depending on the treated patient. Dosing and the introduction of regulated so as to provide sufficient levels of the active fraction or maintain the desired effect. In the calculation may be additional factors, such as severity of disease, age of the organism and the weight or size of the body; diet, time and frequency of administration, combination of drugs, reaction sensitivities, and tolerance/response to therapy. Short-acting pharmaceutical compositions are injected daily or more often, while long-acting pharmaceutical compositions are injected every 2 days or more, once a week or every two weeks, or even less.

The routes of introduction pharmaceuticals are not in order of limitation, local, transdermal, parenteral, gastrointestinal, chrisb anchialine and kresolverentry. Percutaneous introduction is performed by applying a cream, rinse, gel, etc. capable of causing penetration of obmenivatsa specificity of ligand/receptor through the skin. For parenteral routes of introduction are not in order restrictions, electric or direct injection, such as direct injection into a Central venous line, intravenous, intramuscular, intraperitoneal, intradermal or subcutaneous injection. Gastrointestinal routes of introduction are not in the order of limits, ingestion, and rectal administration. To responcibly and resolvehostname routes of introduction is not in order restrictions, inhalation through the mouth or through the nose.

The compositions containing the described obmanivali specificity of ligand/receptor suitable for percutaneous or topical administration, include, but are not in order of limitation, pharmaceutically acceptable suspensions, oils, creams and ointments applied directly to the skin or embedded in a protective carrier such as a transdermal device ("transdermal patch"). Examples of suitable creams, ointments, etc. can be found, for example, in Physician''s Desk Reference. Examples of suitable percutaneous devices are described, for example, in U.S. patent No. 4818540, issued April 4, 1989 Cinena etc.

The compositions containing the described obmanivali specificity of ligand/receptor, prigoda the s for parenteral administration, include, but are not in order of limitation, pharmaceutically acceptable sterile isotonic solutions. Such solutions are not in the order of limits, the saline solution and phosphate-buffered saline for injection into a Central venous line, intravenous, intramuscular, intraperitoneal, intradermal and subcutaneous injection.

The compositions containing the described obmanivali specificity of ligand/receptor, suitable for responcible and resolvehostname introduction, include, but are not in the order limitations, various types of aerosols for inhalation. Devices suitable for their responcible and resolvehostname introduction, are also variants of execution. These devices include, in no particular order restrictions, atomizers and vaporizers. Many forms are already available atomizers and vaporizers can be easily adapted for delivery described herein compounds containing obmanivali specificity of ligand/receptor.

The compositions containing the described obmanivali specificity of ligand/receptor, suitable for gastrointestinal administration, include, but are not in order of limitation, pharmaceutically acceptable powders, pills or liquids for ingestion and suppositories for rectal administration. Because of the ease of use of the gastro-Kish is offered by the introduction, in particular oral, is the preferred implementation. When a pharmaceutical preparation containing obmenival specificity of ligand/receptor obtained, he may be in need of a body for treatment or prevention of pathogenic infection.

Aspects of the invention also includes covering such medical equipment as dentures, implants and instruments. Coatings suitable for use on medical devices can be provided by using a gel or powder containing obmenival specificity of ligand/receptor, or by using a polymer coating, which suspended obmenival specificity of ligand/receptor. Suitable for covering devices are those polymeric materials which are physiologically acceptable and through which can diffuse therapeutically acceptable amount of obmenivalas specificity of ligand/receptor. Suitable polymers include, not in order of limitation, polyurethane, polymethacrylate, polyamide, polyester, polyethylene, polypropylene, polystyrene, polytetrafluoroethylene, polyvinyl chloride, cellulose acetate, silicon elastomers, collagen, silk, etc. Such coatings are described, for example, in U.S. patent No. 4612337.

The following section describes the methods of treatment and prevention of diseases with the help of the described here obmenivatsa specificity of ligand/receptor.

Treatment and prevention of disease using obmenivalas specificity of ligand/receptor

Pharmaceutical preparations containing obmenival specificity of ligand/receptor may be administered to a subject in need of treatment and/or prevention of infection by the pathogen with the receptor. Such needy subjects may include individuals with a risk of contact with the pathogen or individuals already infected with the pathogen. These individuals can be identified using standard clinical or diagnostic methods.

In accordance with one approaches a subject suffering from a bacterial infection, is identified as the subject in need of the agent that inhibits proliferation of a pathogen. Then this subject is provided a therapeutically effective amount of obmenivalas specificity of ligand/receptor. Used in this way obmenival specificity of ligand/receptor contains the domain specificity, interacting with the audience on the bacterial receptor (e.g., extracellular fibrinogen binding protein (Efb), linking the collagen protein, vitronectin binding protein, laminin binding protein, plasminogen binding protein, binding thrombospondin protein aggregation factor A (ClfA), an aggregating factor B (ClfB), tie the state of fibronectin protein, the coagulase and extracellular protein adhesion). Obmenival specificity of ligand/receptor also contains antigenic domain that has an epitope of a pathogen or toxin, preferably, the epitope recognized by antibodies with high titer present need of the subject. It may also be desirable to analyze the needs of the subject antibody with high titer, which recognize antigenic domain, before granting entity obmenivalas specificity of ligand/receptor. This test can be performed by fit or ELISA using immobilized antigenic domain obmenivalas specificity of ligand/receptor, as described above.

In a similar way to a subject in need of the agent, inhibiting a viral infection, can be entered obmenival specificity of ligand/receptor that recognizes a receptor present on a specific etiologic agent. Accordingly, the subject in need of the agent, inhibiting a viral infection, identified by standard clinical or diagnostic procedures. Then need a subject is provided a therapeutically effective amount of obmenivalas specificity of ligand/receptor, which interacts with a receptor present on the virus, the infected individual. As before, it may be desirable in order to determine does the entity sufficient titer of antibodies to interact with the antigenic domain of obmenivalas specificity of ligand/receptor, before the introduction of obmenivalas specificity of ligand/receptor.

Similarly to a subject in need of the agent, inhibiting the proliferation of cancer may be obmenival specificity of ligand/receptor that interacts with a receptor present on a cancer cell. For example, a subject in need of the agent, inhibiting the proliferation of cancer is identified using standard clinical or diagnostic procedures; then requiring the subject is provided a therapeutically effective amount of obmenivalas specificity of ligand/receptor that interacts with a receptor present on cancer cells, infecting the subject. As noted above, it may be desirable to determine whether the subject of sufficient titer or antibody-based test for interaction with antigenic domain obmenivalas specificity of ligand/receptor, prior to the introduction of obmenivalas specificity of ligand/receptor.

Described here obmanivali specificity of ligand/receptor can also be administered to subjects as a prophylactic to prevent the onset of disease. Described here obmenival specificity of ligand/receptor may be virtually any is in the preventive purposes (for example, to prevent bacterial infection, viral infection or cancer). However, it is desirable that the subjects with a high risk of Contracting specific diseases identified and supplied with obmenivalas specificity of ligand/receptor. For subjects with a high risk of Contracting the disease include individuals with a family history of the disease, in the elderly or young, or individuals, often in contact with the pathogen (for example, medical practitioners). Accordingly, subjects with the risk of infection by the pathogen with the receptor are identified, and then supplied prophylactically effective amount of obmenivalas specificity of ligand/receptor.

One of these preventive applications for obmenivatsa specificity of ligand/receptor refers to the planning or application of obmenivalas specificity of ligand/receptor on the medical device or implant. Implantable medical devices serve as foci of infection for certain types of bacteria. Such related devices infection is supported by the tendency of these organisms to adhere to the surface of the device and to colonize it. Therefore, there is a significant need to develop such surfaces, which are less susceptible to reverse biological reactions, which usually is accompanied by the implantation of a medical device.

In accordance with one approach, the medical device is covered with a solution containing obmenival specificity of ligand/receptor. Before implantation of the medical device (for example, the valve prosthesis) may be stored, for example, in the solution of obmenivatsa specificity of ligand/receptor. Medical devices can also be coated with powder or gel containing obmenival specificity of ligand/receptor. For example, gloves, condoms and intrauterine devices may be coated with powder or gel containing obmenival specificity of ligand/receptor that interacts with bacterial or viral receptor. Being implanted in the body, these obmanivali specificity of ligand/receptor provide preventative barrier to infection by the pathogen.

In some embodiments, performing obmenival specificity of ligand/receptor immobilities on the medical device. As described above, the medical device is a base to which may be attached obmenival specificity of ligand/receptor. Immobilization may occur by hydrophobic interactions between obmenivalas specificity of ligand/receptor and medical device, but the preferred method of immobilization of obmenivalas specificity of ligand/receptor is and the medical device is a covalent attachment. For example, the medical device can be produced with a reactive group that interacts with a reactive group located on obmanivali specificity.

In accordance with one embodiment of periodat combined with obmenivalas specificity of ligand/receptor containing 2-aminopyrrolo share, forming an aldehyde-functional obmenival in an aqueous solution having a pH from about 4 to 9 and a temperature of approximately from 0 to 50 degrees Celsius. Then the aldehyde-functional obmenival combined with containing the biomaterial surface of the medical device, which contains a primary amine share to the immobilized obmenival specificity of ligand/receptor on the surface of the substrate through the imine share. Then imine share reacts with regenerating agent, forming an immobilized obmenival specificity of ligand/receptor containing the biomaterial surface through a secondary amine linkage. Other approaches for linking molecules to medical devices (such as described in U.S. patent No. 6017741) can be modified for immobilization described here obmenivalas specificity of ligand/receptor.

Although the invention is described with reference to options and run the examples, you should understand what can be done different is odificatio without departing from the substance of the invention. Accordingly, the invention is limited only by the following claims.

1. Obmenival specificity of ligand/receptor, including

at least one domain specificity, containing the ligand for the receptor, and the ligand is not an antibody or part of it, and

at least one antigenic domain, coupled with the aforementioned domain specificity, in which the aforementioned antigenic domain contains the epitope of a pathogen or toxin.

2. Obmenival specificity of ligand/receptor according to claim 1, in which the mentioned domain specificity contains at least three consecutive amino acids of a peptide selected from the group consisting of extracellular matrix protein, a ligand for receptors on the virus and the ligand for a receptor on a cancer cell.

3. Obmenival specificity of ligand/receptor according to claim 2, in which the said peptide is an extracellular matrix protein selected from the group consisting of fibrinogen, collagen, vitronectin, laminin, plasminogen, thrombospondin and fibronectin.

4. Obmenival specificity of ligand/receptor according to claim 2, in which the said peptide is a ligand for a receptor on a virus selected from the group consisting of glycopro thein T4 and envelope protein of hepatitis C.

5. Obmenival specificity of ligand/receptor according to claim 2, in which the said peptide is a ligand for a receptor on a cancer cell selected from the group consisting of a ligand for HER-2/neu and ligand for integranova receptor.

6. Obmenival specificity of ligand/receptor according to claim 1, in which the mentioned domain specificity contains at least one sequence selected from the group consisting of SEQ.ID.NO.:1, SEQ.ID.NO.:2, SEQ.ID.NO.:3, SEQ.ID.NO.:4, SEQ.ID.NO.:5, SEQ.ID.NO.:6, SEQ.ID.NO.:7, SEQ.ID.NO.:8, SEQ.ID.NO.:9, SEQ.ID.NO.:10, SEQ.ID.NO.:11, SEQ.ID.NO.:12, SEQ.ID.NO.:13, SEQ.ID.NO.:14, SEQ.ID.NO.:15, SEQ.ID.NO.:16, SEQ.ID.NO.:17, SEQ.ID.NO.:18, SEQ.ID.NO.:19, SEQ.ID.NO.:20, SEQ.ID.NO.:21, SEQ.ID.NO.:22, SEQ.ID.NO.:23, SEQ.ID.NO.:24, SEQ.ID.NO.:25, SEQ.ID.NO.:26, SEQ.ID.NO.:27, SEQ.ID.NO.:28, SEQ.ID.NO.:29, SEQ.ID.NO.:30, SEQ.ID.NO.:31, SEQ.ID.NO.:32, SEQ.ID.NO.:33, SEQ.ID.NO.:34, SEQ.ID.NO.:35, SEQ.ID.NO.:36, SEQ.ID.NO.:37, SEQ.ID.NO.:38, SEQ.ID.NO.:39, SEQ.ID.NO.:40, SEQ.ID.NO.:41, SEQ.ID.NO.:42 and SEQ.ID.NO.:124.

7. Obmenival specificity of ligand/receptor according to claim 3, in which the aforementioned extracellular matrix protein comprises at least 3 amino acids of the alpha-chain of fibrinogen.

8. Obmenival specificity of ligand/receptor according to claim 1 in which the said ligand contains a sequence arginine-glycine-aspartate (RGD).

9. Obmenival specificity of ligand/receptor according to claim 1 in which the said receptor is located in the pathogen.

10. Obmenival specificity of ligand/receptor according to claim 1 in which the said receptor is bacterial is desioner receptor.

11. Obmenival specificity of ligand/receptor of claim 10, in which the mentioned bacterial adhesion receptor is selected from the group consisting of extracellular fibrinogen binding protein (Efb), linking the collagen binding protein vitronectin binding protein laminin binding protein a plasminogen binding protein of thrombospondin protein aggregation factor A (ClfA), aggregating factor B (ClfB), fibronectin binding protein, coagulase and protein extracellular adhesion.

12. Obmenival specificity of ligand/receptor according to claim 1, in which the aforementioned antigenic domain contains at least three amino acid peptide selected from the group consisting of a protein of herpes simplex virus, squirrel hepatitis b virus, the protein of TT virus and protein of poliovirus.

13. Obmenival specificity of ligand/receptor indicated in paragraph 12, in which the aforementioned antigenic domain is a protein of herpes simplex virus containing at least one sequence selected from the group consisting of SEQ.ID.NO.:53 and SEQ.ID.NO.:54.

14. Obmenival specificity of ligand/receptor indicated in paragraph 12, in which the aforementioned antigenic domain is a protein of hepatitis b virus containing at least one sequence selected from the group consisting of SEQ.ID.NO.:49, SEQ.ID.NO.:50, SEQ.ID.NO.:52 and SEQ.ID.NO.:59.

15. Obmenival specificness and ligand/receptor indicated in paragraph 12, in which mentioned antigenic domain is a protein of TT virus containing at least one sequence selected from the group consisting of SEQ.ID.NO.:43, SEQ.ID.NO.:44, SEQ.ID.NO.:45, SEQ.ID.NO.:46, SEQ.ID.NO.:47, SEQ.ID.NO.:55, SEQ.ID.NO.:56, SEQ.ID.NO.:57 and SEQ.ID.NO.:58.

16. Obmenival specificity of ligand/receptor indicated in paragraph 12, in which the aforementioned antigenic domain is a protein of poliovirus containing at least one sequence selected from the group consisting of SEQ.ID.NO.:48 and SEQ.ID.NO.:51.

17. Obmenival specificity of ligand/receptor according to claim 1, in which the aforementioned antigenic domain interacts with the antibody with high titer.

18. Obmenival specificity of ligand/receptor on 17, in which the aforementioned antigenic domain specifically binds to an antibody present in the serum of the animal, which was diluted to approximately 1:100-1:1000 or greater.

19. Obmenival specificity of ligand/receptor according to claim 1, in which the sequence mentioned obmenivalas specificity of ligand/receptor is selected from the group consisting of SEQ.ID.NO.:60, SEQ.ID.NO.:61, SEQ.ID.NO.:62, SEQ.ID.NO.:63, SEQ.ID.NO.:64, SEQ.ID.NO.:65, SEQ.ID.NO.:66, SEQ.ID.NO.:67, SEQ.ID.NO.:68, SEQ.ID.NO.:69, SEQ.ID.NO.:70, SEQ.ID.NO.:71, SEQ.ID.NO.:72, SEQ.ID.NO.:73, SEQ.ID.NO.:74, SEQ.ID.NO.:75, SEQ.ID.NO.:76, SEQ.ID.NO.:77, SEQ.ID.NO.:78, SEQ.ID.NO.:79, SEQ.ID.NO.:80, SEQ.ID.NO.:81, SEQ.ID.NO.:82, SEQ.ID.NO.:83, SEQ.ID.NO.:84, SEQ.ID.NO.:85, SEQ.ID.NO.:86, SEQ.ID.NO.:87, SEQ.ID.NO.:88, SEQ.ID.NO.:89, SEQ.ID.NO.:90, SEQ.ID.NO.:91, SEQ.ID.NO.:92,SEQ.ID.NO.:93, SEQ.ID.NO.:94, SEQ.ID.NO.:95, SEQ.ID.NO.:96, SEQ.ID.NO.:97, SEQ.ID.NO.:98, SEQ.ID.NO.:99, SEQ.ID.NO.:100, SEQ.ID.NO.:101, SEQ.ID.NO.:102, SEQ.ID.NO.:103, SEQ.ID.NO.:104, SEQ.ID.NO.:105, SEQ.ID.NO.:137 and SEQ.ID.NO.:142.

20. Obmenival specificity of ligand/receptor according to claim 1, in which the mentioned domain specificity has a length of at least 3 amino acids.

21. Obmenival specificity of ligand/receptor according to claim 1, in which the mentioned domain specificity has a length of at least 8 amino acids.

22. Obmenival specificity of ligand/receptor according to claim 1, in which the mentioned domain specificity has a length of at least 20 amino acids.

23. Obmenival specificity of ligand/receptor according to claim 1, in which the mentioned domain specificity contains a ligand for bacterial adhesion receptor.

24. Obmenival specificity of ligand/receptor according to claim 1, in which the mentioned domain specificity contains at least 3 consecutive amino acids of fibrinogen.

25. Obmenival specificity of ligand/receptor for item 23, in which the mentioned bacterial adhesion receptor is an adhesion receptor Staphylococcus.

26. Obmenival specificity of ligand/receptor according to claim 9, in which the said pathogen is Staphylococcus (Staphylococcus).

27. The glycosylated obmenival specificity of ligand/receptor containing

at least one domain specificness is, containing at least 3 consecutive amino acids of fibrinogen, which are associated with bacterial adhesion receptor; and

at least one antigenic domain, coupled with the aforementioned domain specificity, and mentioned antigenic domain contains the epitope of a pathogen or toxin and these obmenival specificity of ligand/receptor glycolysation.

28. Obmenival specificity of ligand/receptor containing saccharide and at least one domain specificity, containing at least 3 consecutive amino acids of fibrinogen, which bind bacterial adhesion receptor, coupled with antigenic domain that contains the epitope of a pathogen or toxin.

29. Domain specificity, containing the ligand for the receptor, in which the said ligand is selected from the group consisting of SEQ.ID.NO.:106, SEQ.ID.NO.:107, SEQ.ID.NO.:108, SEQ.ID.NO.:109, SEQ.ID.NO.:110, SEQ.ID.NO.:111, SEQ.ID.NO.:115, SEQ.ID.NO.:116, SEQ.ID.NO.:119, SEQ.ID.NO.:121, SEQ.ID.NO.:122, SEQ.ID.NO.:123, SEQ.ID.NO.:124, SEQ.ID.NO.:125, SEQ.ID.NO.:127, SEQ.ID.NO.:128, SEQ.ID.NO.:129, SEQ.ID.NO.:130, SEQ.ID.NO.:131, SEQ.ID.NO.:132, SEQ.ID.NO.:133, SEQ.ID.NO.:134 that is associated with the polysaccharide.

30. Domain specificity of ligand/receptor containing the domain specificity of containing the ligand for bacterial adhesion receptors and antigenic domain, connected with the domain specificity, in which antigenic domain contains the epitope of the bacteria the branches of the pathogen.



 

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FIELD: molecular biology, biochemistry, medicine, oncology.

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FIELD: genetic engineering, medicine.

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FIELD: medicine.

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FIELD: biotechnology, molecular biology, medicine.

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FIELD: biotechnology, peptides, genetic engineering.

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FIELD: genetic engineering, pharmaceutical and medical-biological industry.

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FIELD: biotechnology, molecular biology, medicine.

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EFFECT: valuable biological, medicinal and veterinary properties of polypeptide.

23 cl, 71 dwg, 12 tbl, 17 ex

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FIELD: genetic engineering, pharmaceutical and medical-biological industry.

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23 cl, 67 dwg, 1 tbl, 35 ex

FIELD: biotechnology, microbiology, genetic engineering.

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EFFECT: valuable biological and medicinal properties of polypeptide.

3 dwg, 4 ex

FIELD: medicine, oncology, biochemistry.

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EFFECT: valuable medicinal properties of protein complexes.

13 cl, 40 dwg, 18 ex

FIELD: biotechnology, medicine.

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EFFECT: valuable properties of new cytokine.

18 cl, 5 tbl, 1 dwg, 55 ex

FIELD: genetic and tissue engineering, biotechnology, medicine, agriculture.

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EFFECT: improved preparing method, valuable medicinal properties of vector, improved genetic modification.

7 cl, 12 dwg, 4 tbl, 16 ex

The invention relates to genetic engineering and can be used for therapeutic purposes, in particular in the treatment of neoplastic processes

FIELD: medicine; biology.

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EFFECT: enhanced effectiveness in treating pathological states with new blood vessel formation being suppressed.

7 cl, 8 dwg

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