title: Monoclonal antibodies requiring coating buffer with low pH for efficient antigen capture in sandwich ELISA: the rarities or practically important phenomena?
abstract: This article reexamines some opinions concerning pH requirements for optimal immobilization of monoclonal antibodies (mAbs) by passive adsorption in antigen capture ELISA. It was discovered that substitution of "classical" sodium phosphate (pH 7.5) and carbonate (pH 9.5) coating solutions by acid (pH 2.8) buffers maximized antigen capture 4 out of 10 different tested anti-HBsAg mAbs, resulting in a 1.5-2.5 increase of binding curve coefficients. By measuring both mAbs amounts and functionality, the enhancement effect was attributed to the better preservation of solid phase antibodies activity.
authors.affiliation: Mechnikov Research Institute of Vaccines and Sera, RAMS, O. Andzhaparidze Virology Unit, Moscow, Russia. dmitrievda@mail.ru
authors.forename: Alexander D
authors.initials: AD
authors.lastname: Dmitriev
authors.forename: Julia N
authors.initials: JN
authors.lastname: Tarakanova
authors.forename: Dinora A
authors.initials: DA
authors.lastname: Yakovleva
authors.forename: Dmitriy A
authors.initials: DA
authors.lastname: Dmitriev
authors.forename: Olga V
authors.initials: OV
authors.lastname: Phartooshnaya
authors.forename: Galina I
authors.initials: GI
authors.lastname: Kolyaskina
authors.forename: Yulia S
authors.initials: YS
authors.lastname: Massino
authors.forename: Olga V
authors.initials: OV
authors.lastname: Borisova
authors.forename: Olga L
authors.initials: OL
authors.lastname: Segal
authors.forename: Maria B
authors.initials: MB
authors.lastname: Smirnova
authors.forename: Tatiana I
authors.initials: TI
authors.lastname: Ulanova
authors.forename: Viacheslav F
authors.initials: VF
authors.lastname: Lavrov
mesh_terms: D000327:Adsorption; D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000276:immunology*; D000937:Antigen-Antibody Reactions; D000941:Antigens / Q000302:isolation & purification*; D002021:Buffers; D004797:Enzyme-Linked Immunosorbent Assay / Q000295:instrumentation / Q000379:methods*; D006514:Hepatitis B Surface Antigens / Q000276:immunology*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007118:Immunoassay / Q000379:methods; D011817:Rabbits
chemical_list: D000911:Antibodies, Monoclonal; D000941:Antigens; D002021:Buffers; D006514:Hepatitis B Surface Antigens