Compositions containing secretor-like immunoglobulins
SUBSTANCE: method for in vitro preparation of a composition comprising a secretor-like immunoglobulin comprising the steps of production of a protein composition isolated from blood containing an immunoglobulin containing a J-chain in a crude form and mixing of the composition of step (a) with a secretory component. A composition for use in medicine comprising secretor-like IgA and/or secretor-like IgM or a combination thereof obtained by the described method is provided.
EFFECT: increased efficiency of composition application.
17 cl, 8 dwg, 5 ex
SUBSTANCE: invention relates to biochemistry. Described are antibodies and fragments thereof, having high affinity for human α-synuclein protofibrils and low binding of α-synuclein monomers. Disclosed are compositions which contain the described antibody or a fragment thereof, methods of detecting α-synuclein protofibrils using the described antibody or fragment thereof. In other versions, the present invention relates to methods of preventing, slowing down the development or treating a neurodegenerative disorder with α-synuclein pathology, where said methods include administering the described antibody or fragment thereof. The invention also relates to use of said antibody or fragment thereof to obtain a pharmaceutical composition for treating a neurodegenerative disorder with α-synuclein pathology.
EFFECT: invention is used for diagnosis or monitoring of the development of a neurodegenerative disorder with α-synuclein pathology, and in methods of reducing or inhibiting α-synuclein aggregation by administering said antibody or fragment thereof.
31 cl, 9 dwg, 4 tbl, 10 ex
SUBSTANCE: method comprises immunoaffinity chromatography using mini-antibodies a-hLF-1 and a-hLF-4, which amino acid sequences are presented as SEQ ID NO:1 and SEQ ID NO:2. The invention may be used to obtain highly purified fraction of human lactoferrin.
EFFECT: invention enables to carry out with high efficiency the separation of proteins of lactoferrin of human and goat consisting in milk, using the single-domain mini-antibodies.
6 dwg, 2 ex
SUBSTANCE: invention refers to biotechnology and immunology. The method provides administering an antibody which binds to an advanced glycation end product. What is also described is using this antibody for producing a therapeutic agent. The invention can be used in medicine.
EFFECT: disclosed is a method for promoting the regenerative processes in a tissue culture or a cell culture.
12 cl, 1 dwg, 2 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: present invention refers to biotechnology and immunology. What is presented is a hybridoma cell line FP12H3-C2 deposited under No. DSM ACC2750 producing the anti-beta-amyloid antibody. Presented are methods for preparing an antibody, including a humanised antibody with using a hybridoma line according to the invention, as well as diagnostic techniques for a beta-amyloid associated disease or condition in a patient or a liability to such disease or condition, a method for determining a degree of the tissue involvement into amyloidogenic plaques, a method for monitoring minimal residual signs of the disease into a patient following treating with the antibody or its active fragment, a method for prediction of sensitivity in the patient treated with the antibody or its active fragment.
EFFECT: present invention can find further application in diagnosing and therapy of beta-amyloid related diseases, such as Alzheimer disease.
9 cl, 4 dwg, 5 tbl, 17 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: present invention refers to biotechnology and represents anti-nerve growth factor (NGF) antibodies. The present invention also discloses a pharmaceutical composition for relieving pain associated with a disease or a condition, wherein pain progression or persistence is mediated by NGF, containing the above antibodies, as well as a kit for treating a HGF-related disease, such as e.g. osteoarthritis, nucleic acids coding a heavy or light chain of the antibody, an expression vector, a host cell for preparing the above antibodies, a method for expressing the above anti-NGF antibodies, as well as using the above antibodies in managing pain and for preparing a therapeutic agent for managing pain associated with the disease or condition, wherein pain progression or persistence is mediated by NGF.
EFFECT: present invention enables producing the anti-NGF antibodies characterised by high stability in vivo.
16 cl, 7 dwg, 13 tbl, 8 ex
SUBSTANCE: invention refers to immunology. Presented are anti-Dickkopf 1 (anti-Dkk-1) antibodies and their functional fragments specified among the antibodies: 1) containing CDR1 VH containing the amino acid sequence SSYAIS, SYAIS or GFTFSSY; CDR2 VH containing the amino acid sequence SVSGTGLGFGTYYPDSVKG or SVSGTGLGFGTY; and CDR3 VH, containing the amino acid sequence TSLENYAFDY or SLENYAFDY; and CDR1 VL containing the amino acid sequence RASESVDDFGISFIN; CDR2 VL containing the amino acid sequence AGSKQGS; and CDR3 VL containing the amino acid sequence QQLKEVPPT; and 2) the antibodies disclosed in Table 4 presented in the application materials. Described are: nucleic acids coding the above antibodies or their functional fragments; expression vectors containing the above nucleic acids; and cells used for expression of the above antibodies or their functional fragments and containing the above expression vectors. Presented is a method for producing the antibody or its functional fragment involving the stage of culturing the above expression cell. Disclosed is a composition possessing Dkk-1 binding activity, containing the antibody or its functional fragment in a therapeutically effective amount and a pharmaceutically acceptable excipient, thinner or carrier.
EFFECT: invention enables extending the range of products for treating the diseases associated with Dkk-1 and LRP5/6 excessive reaction, which cause Wnt activation.
14 cl, 14 dwg, 14 tbl, 6 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to biotechnology. There are described pharmaceutical compositions for treating ophthalmic diseases associated with pathological abnormalities/changes of visual tissues, especially associated with amyloid-beta related pathological abnormalities/changes of the visual tissues, containing antibodies specifically recognising and binding specific epitopes from the range of β-amyloid proteins. There are also presented methods for reducing the load/plaque count in the retinal gangliocyte layer in a mammal suffering from an analogous ophthalmic disease. Besides, there are presented a method for reducing the total amount of soluble amyloid-β in the mammalian retinal gangliocyte layer, a method for preventing manifestations of the analogous ophthalmic disease, a method of treating or relieving the manifestations of the analogous ophthalmic disease. A method for maintaining or reducing an ocular tension in an animal suffering from the analogous ophthalmic disease. All the methods involve administering the described composition.
EFFECT: invention extends the methods and compositions for diagnosing and treating the amyloid diseases that represent a group of amyloid protein related diseases and disorders, eg Alzheimer disease.
55 cl, 20 dwg, 7 tbl, 18 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to biotechnology and immunology. What is described is a pharmaceutical composition used for treating and/or preventing pathological bone metabolism and containing this antibody. The invention can be used in medicine.
EFFECT: antibody and its functional fragment specifically recognising human Siglec-15 and possessing the osteoclast inhibitory activity are described.
73 cl, 57 dwg, 4 tbl, 33 ex
SUBSTANCE: group of inventions refers to medicine, namely to ophthalmology, and can be used for treating ocular diseases associated with amyloid-beta related pathological abnormalities/changes in the visual system tissues. That is ensured by administering a pharmaceutical composition, which contains a therapeutically effective amount of a humanised antibody or antigen-binding fragment, wherein the humanised antibody or its fragment is able to bind amyloid-beta. Presented are preventing, treating or relieving symptoms of an ocular disease, reducing the plaque load of retinal ganglion cells, diagnosing the ocular disease and diagnosing a predisposition to the ocular disease, prolonging the patient's sensitivity when treating with the pharmaceutical composition for treating the ocular disease.
EFFECT: group of inventions provides the effective treating of the above ocular pathology by using the composition containing the high-specific antibodies, which specifically recognise and bind to specific epitopes of various β-amyloid proteins.
20 cl, 18 dwg, 9 tbl, 18 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to biochemistry. What is described is an anti-Tau phospho Serine 422 (pS422) antibody for treating taupathy, characterised by the fact that it specifically binds to a phosphorylated Tau fragment of the sequence SEQ ID NO:9 and to Tau pS422, and binds to other than Tau and phosphorylated MSAC fragment of the sequence SEQ ID NO:17. What is presented is the pharmaceutical composition for treating taupathy, as well as a method of treating taupathy. What is presented is a method for preparing the above antibody.
EFFECT: invention extends the range of products for treating taupathy.
21 cl, 11 dwg, 7 tbl, 12 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: presented invention refers to immunology. There are disclosed versions of a dimer compound for forming a multimer capable to reproduce the effector function of aggregated IgG with identical monomers. Each monomer of the dimer comprises: a monomer of IgG2 link region or a monomer of isoleucine zipper, dimerising each of which forms a multimerising region, and at least Fc-domain monomer containing a link region, CH2 domain and CH3 domain of IgG1. What is described is a multimer compound capable to reproduce the effector function of aggregated IgG and containing two or more dimers. There are disclosed a method for changing the immune response using the dimer or multimer, as well as a multimer-based method of treating an inflammatory disease.
EFFECT: using the invention provides the new compounds capable to bind at least one FcR specified in a group consisting of: FcγRI, FcγRII, FcγRIII, FcγRIV, or their non-human version that can find application in medicine for IVIG substitution for treating a wide range of diseases, including the inflammatory and autoimmune diseases.
7 cl, 25 dwg, 5 tbl, 25 ex
SUBSTANCE: invention relates to biochemistry. Described is a bispecific antigen-binding protein which is heterodimeric with respect to binding protein A. The protein contains a first polypeptide comprising, from N-terminal to C-terminal, a first epitope-binding region that selectively binds a first epitope, an immunoglobulin constant region that comprises a first CH3 region of a human IgG selected from IgG1, IgG2, and IgG4, where the first CH3 region binds with protein A, and a second polypeptide comprising, from N-terminal to C-terminal, a second epitope-binding region that selectively binds a second epitope, an immunoglobulin constant region that comprises a second CH3 region of a human IgG selected from IgG1, IgG2, and IgG4, where the second CH3 region comprises a modification that reduces or prevents binding of the second CH3 domain to protein A.
EFFECT: invention enables rapid isolation of protein A.
9 cl, 10 dwg, 3 tbl, 12 ex
SUBSTANCE: disclosed is a method of purifying antibodies, including human CD20 and VEGF binding antibodies, from a composition which contains an antibody and at least one impurity compound, said method comprising the following steps: (a) loading the composition on a cation-exchange material, where said composition has first pH value from 4.0 to 6.0; (b) washing the cation-exchange material with a first washing buffer at pH higher than that of composition (a), where pH of the first washing buffer ranges from 6.8 to 9.0; (c) washing the cation-exchange material with a second washing buffer at pH lower than that of the first washing buffer; where the second washing buffer has conductivity from 0.5 to 3.0 mS/cm and pH from 5.0 to 6.0; and (d) elution of the antibodies from the cation-exchange material with an elution buffer at conductivity which is at least 2 mS/cm higher than that of the second washing buffer, where pH of the second washing buffer and pH of the elution buffer are approximately equal, and where pH of the elution buffer ranges from 5.0 to 6.0. Described is a conjugation method, which involves conjugating the purified product obtained using said method with a heterologous molecule. Also disclosed is a method of producing a pharmaceutical composition, which involves combining said purified product with a pharmaceutically acceptable carrier.
EFFECT: invention improves purification of antibodies used for treatment.
16 cl, 3 dwg, 4 tbl, 2 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention discloses a method for producing high-affinity polyclonal antibodies. The method involves the stages as follows: a) application of an antibody solution on an affine sorbent with an immobilised antibody at an excess molar quantity of antigen antigen-specific antibodies; b) removal of proteins and low-affinity antibodies; c) removal of the bound medium-affinity antibodies by means of a moderately chaotropic agent, such as LiCl, MgCl2, glycine, d) elution of the high-affinity antibodies by means of the chaotropic agent, e) removal of the latter and f) if needed, removal of antibody aggregates. The method provides producing the antibodies affinity and specificity of which substantially exceeds those of the polyclonal antibodies included in the state of the art.
EFFECT: use of the antibodies prepared by the method according to the invention enables prominently increasing sensitivity and specificity of immunoassay techniques.
13 cl, 5 dwg, 1 tbl, 4 ex
SUBSTANCE: invention relates to liquid chromatography. Disclosed is a method for chromatographic extraction of immunoglobulin, involving dissolution of a blood plasma protein fraction in a buffer solution, said blood plasma protein fraction being a residue A of blood plasma alcohol fractionation via a Cohn process. The obtained solution undergoes preliminary purification in two series-connected columns filled with a hydrophobic sorbent and anionite, respectively, and then passing a buffer solution through said two columns. After collecting the purified liquid fraction, which contains immunoglobulin, it is taken for solvent-detergent viral inactivation and then for chromatographic purification, which is carried out in a system of three series-connected columns filled with anionite, hydrophobic sorbent and cationite, respectively. The immunoglobulin from the column filled with cationite is eluted and columns with the anionite and hydrophobic sorbent are taken for regeneration.
EFFECT: invention enables to extract immunoglobulin from crude material - a residue A, obtained from blood plasma alcohol fractionation via a Cohn process, with high output and purity of the end product.
8 cl, 2 ex
SUBSTANCE: method of monomer antibody recovery from a compound containing large molecular weight antibody aggregates provides contacting of the compound with hydroxyapatite resin and elution of purified antibody from resin. There are also disclosed versions of such method providing protein A affinity chromatography, ion-exchange chromatography and hydroxyapatite chromatography.
EFFECT: methods allow producing a compound with the lower content of large molecular weight antibody aggregates.
23 cl, 4 dwg, 14 tbl, 11 ex
FIELD: chemistry; biochemistry.
SUBSTANCE: invention relates to biotechnology, specifically to obtaining a human immunoglobulin based preparation, and can be used in medicine. The preparation is obtained via purification of class G, A and M immunoglobulins isolated from the blood of HIV infected patients through affinity chromatography on a column with integrase-sepharose.
EFFECT: invention enables to obtain class G, A and M immunoglobulins isolated from the blood of HIV infected patients, capable of selectively splitting HIV integrase only.
7 dwg, 4 ex
SUBSTANCE: invention refers to virus inactivation in production of immunoglobulins, particularly of G fraction. The method virus inactivation for production of G fraction immunoglobulin includes purification of dissolved immunoglobulin recovered by Kohn's spirit fractionation. Purified dissolved immunoglobulin is prepared with a solvent detergent mixture which is acetate buffer solution 0.05 M at pH 5.5, containing 1 wt % tri-n-butylphosphate and 1 wt % polysorbate 80. Preparation represents stirring of the mixture within 12-16 hours followed with dilution with acetate buffer solution 0.05 M at pH 5.5 containing 1 wt % sodium octanoate, sodium chloride 0.15 M and propylene glycol concentrated 0.2 g/l. Prepared immunoglobulin is immobilised on sulphoprolylcation sorbent with grain size 50 mcm and sorption capacity 55 mg/cm3, and two-stage washed by column chromatography followed with elution. The first stage of washing implies acetate buffer solution 0.05 M at pH 5.5 containing 1 wt % sodium octanoate, sodium chloride 0.15 M and propylene glycol concentrated 0.2 g/l. The second stage of washing applies acetate buffer solution 0.05 M at pH 5.5. At the first stage the product is washed with volume related to 5 volumes of sorbent, while at the second stage washing is performed with volume related to three volumes of sorbent.
EFFECT: invention provides high-effective virus inactivation that improves viral safety of immunoglobulin preparations.
3 cl, 7 ex
SUBSTANCE: invention refers to medicine, namely to manufacturing of medical products. There is disclosed method for producing immunoglobulin G that involves ethanol fractionation of donor plasma to make sediment B and purification of prepared immunoglobulin. The sediment B is dissolved in 0.9% saline pH 5.15 and mixed with acetate buffer solution 2 M and 53% ethanol, and centrifugated. Centrifugate is mixed with sodium hydrocarbonate at solution pH 5.5. After centrifugation, the centrifugate is purified, mixed with acetate buffer solution at pH 5.4, 96% (vol/vol) ethanol and sodium bicarbonate at pH 7.2, centrifugated at minus (10-12)°C. The recovered sediment is dissolved in acetate buffer solution 0.05 M at pH 5.5, added with solvent detergent mixture containing acetate buffer solution 0.05 M at pH 5.5, and 1 wt % tri-n-butylphosphate and 1 wt % polysorbate 80. The mixture is stirred, then diluted with acetate buffer solution 0.05 M at pH 5.5 containing 1 wt % sodium octanoate, sodium chloride 0.15 M and propylene glycol concentrated 0.2 g/l. Immunoglobulin processed in such a way is immobilised and two-stage washed on sulphoprolylcation sorbent by column chromatography followed with elution, ultrafiltration and sterilisation filtration.
EFFECT: invention provides high-degree purification of immunoglobulin G, absence of viruses and stability of ready preparation.
3 cl, 2 ex
SUBSTANCE: invention concerns medicine and biotechnology, and method of obtaining immunoglobulin medicines. Method of obtaining immunoglobulin medicines involves a sequential processing of alcohol sediment B (Cohn III fraction) by tenfold volume of 0.9% sodium chloride solution and by chloroform in 0.1%-3.0% end concentration, centrifugation, addition of copper sulfate to centrifugate at pH 6-8, sediment removal by centrifugation, and further extraction of target product by ultrafiltration method in the presence of complex-forming compounds, product stabilisation, clearing and stabilising filtration. Copper sulfate is taken in 0.003-0.03% concentration.
EFFECT: increased content of separated immunoglobulin A fraction, improved product quality.
SUBSTANCE: invention relates to the field of obtaining and separation of single-domain molecules (SDAB). Described is a method of the separation or purification of the SDAB molecule, which represents a trivalent molecule of a ATN-103 nanobody, targeting TNFα and HAS, from a mixture, containing the said SDAB molecule and one or more polluting substances. The mixture is brought in contact with a cation-exchange carrier under conditions, which make it possible for the SDAB molecule to bind with the carrier or be absorbed on the carrier. One or more polluting substances are removed and SDAB is selectively eluted from the carrier. The conductivity of a conditioning medium (CM), used for the carrier loading, constitutes from approximately 12 to 9 mS/cm and pH under conditions of loading is corrected to a value from 4.0 to 4.3. The buffer for elution corresponds to approximately 50 mM of sodium chloride or less and has pH from approximately 5.5 to 7.2. Disclosed is a method or a process of obtaining recombinant SDAB of ATN-103. A host-cell is supported in the conditions at which recombinant ATN-103 SDAB is expressed. The mixture of molecule SDAB and one or more polluting substances is obtained. ATN-103 SDAB is purified or separated with the application of cation-exchange chromatography, as said above.
EFFECT: application of the invention provides new methods of the separation or purification of the nanobody, which can be applied in obtaining the ATN-103 nanobody.
19 cl, 4 dwg, 6 ex