Derivation and application of monoclonal antibodies recognizing several epitopes on bovine serum albumin.
Three (AB-3, AB-4 and AB-6) monoclonal antibodies (mAb) to bovine serum albumin (BSA) were derived and characterized for their physicochemical and immunological properties. AB-3 recognized an epitope distinct from epitopes recognized by AB-4 and AB-6 as determined by binding inhibition assay. AB-4 and AB-6 mAbs recognized similar but not identical epitopes on BSA. Based on the antigenic specificity, we applied these mAbs to quantitative analysis of BSA in medium and to depletion of BSA from culture medium containing fetal calf serum (FCS). For quantitative analysis, we employed a sandwich enzyme-linked immunosorbent assay (ELISA) using biotinylated AB-3, solid-phase of AB-6 and an avidin-biotin-peroxidase complex system. This assay was highly sensitive and quantitative in the range of BSA concentration at 10 to 1,500 ng/ml. To deplete BSA from medium, we prepared affinity-gel coupled to AB-6. Repeated treatment of FCS-containing medium with the affinity-gel efficiently depleted BSA from the medium. The depletion capacity was 0.74 to 1.0 moles of BSA/mole of coupled mAb.
Wang
J
J
Department of Hygienic Chemistry, Tohoku University, Sendai.
Masuko
T
T
Ueno
H
H
Hojo
H
H
Hashimoto
Y
Y
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology; D002417:Cattle; D002846:Chromatography, Affinity; D003470:Culture Media / Q000032:analysis; D016895:Culture Media, Serum-Free / Q000032:analysis; D004285:Dogs; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology*; D006801:Humans; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D051381:Rats; D012680:Sensitivity and Specificity; D012710:Serum Albumin, Bovine / Q000276:immunology*
D000911:Antibodies, Monoclonal; D003470:Culture Media; D016895:Culture Media, Serum-Free; D000939:Epitopes; D012710:Serum Albumin, Bovine