title: [Effect of conditions of monoclonal antibody adsorption on antigen-binding activity].
abstract: The dependence of the antigen-binding activity of immobilized antibodies on pH of a saturating buffer has been investigated. We analyzed 28 monoclonal antibodies (MCAs) produced by various hybridomas to three virus antigens, i.e., the nuclear p23 protein of hepatitis C virus (C core protein p23), p24 protein of HIV 1, and the surface antigen of hepatitis B virus (HBsAg). Antibodies were adsorbed on the surfaces of immune plates in acidic (pH 2.8), neutral (pH 7.5), and alkaline (pH 9.5) buffers. The binding of labeled antigens, i.e., biotinylated or conjugated with horseradish peroxidase, with immobilized antigens was tested. It was shown that 10 out of 28 analyzed MCAs (36%) considerably better preserved their antigen-binding activity if their passive adsorption was carried out on the surface of polystyrene plates in an acidic buffer (pH 2.8). This approach allowed constructing a highly sensitive sandwich method for HBsAg assay with a minimal reliably determined antigen concentration of 0.013-0.017 ng/ml. The described approach may be recommended for the optimization of sandwich methods and solid-phase competitive methods.
authors.forename: Iu N
authors.initials: IuN
authors.lastname: Tarakanova
authors.forename: D A
authors.initials: DA
authors.lastname: Dmitriev
authors.forename: Iu S
authors.initials: IuS
authors.lastname: Massino
authors.forename: M B
authors.initials: MB
authors.lastname: Smirnova
authors.forename: O L
authors.initials: OL
authors.lastname: Segal
authors.forename: O V
authors.initials: OV
authors.lastname: Fartushnaia
authors.forename: D A
authors.initials: DA
authors.lastname: Iakovleva
authors.forename: G I
authors.initials: GI
authors.lastname: Koliaskina
authors.forename: V F
authors.initials: VF
authors.lastname: Lavrov
authors.forename: A D
authors.initials: AD
authors.lastname: Dmitriev
mesh_terms: D000327:Adsorption; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000378:metabolism*; D000941:Antigens / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D016655:HIV Core Protein p24 / Q000276:immunology / Q000378:metabolism; D006514:Hepatitis B Surface Antigens / Q000276:immunology / Q000378:metabolism; D018936:Hepatitis C Antigens / Q000276:immunology / Q000378:metabolism; D006735:Horseradish Peroxidase / Q000378:metabolism; D006825:Hybridomas; D006863:Hydrogen-Ion Concentration; D011137:Polystyrenes; D014758:Viral Core Proteins / Q000276:immunology / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D000941:Antigens; D016655:HIV Core Protein p24; D006514:Hepatitis B Surface Antigens; D018936:Hepatitis C Antigens; D011137:Polystyrenes; D014758:Viral Core Proteins; C514799:p24 protein, Human Immunodeficiency Virus Type 1; D006735:Horseradish Peroxidase