Monoclonal antibodies against human alpha-fetoprotein. Exploitation of an unusual calcium-dependent interaction with the antigen for analytical and preparative purposes

• Published in: Journal of immunological methods Vol. 111; no. 1; p. 67
• Main Authors: Stefanová, I, Horejsí, V, Kristofová, H, Angelisová, P, Zizkovský, V, Hilgert, I
• Format: Journal Article
• Language: English
• Published: Netherlands 28-06-1988
• Subjects: alpha-Fetoproteins - analysis; alpha-Fetoproteins - immunology; Animals; Antibodies, Monoclonal - biosynthesis; Antibodies, Monoclonal - immunology; Antigen-Antibody Reactions - drug effects; Binding, Competitive; Chelating Agents - pharmacology; Collodion; Electrophoresis, Polyacrylamide Gel; Enzyme-Linked Immunosorbent Assay; Epitopes - immunology; Female; Humans; Hybridomas - metabolism; Indicators and Reagents; Mice; Mice, Inbred BALB C; Radioimmunoassay; Structure-Activity Relationship
• ISSN: 0022-1759
• DOI: 10.1016/0022-1759(88)90060-9

Seven murine hybridomas were established producing IgG1 monoclonal antibodies (mAbs) against three different epitopes of human alpha-fetoprotein (alpha FP). The interaction of two of these mAbs (AFP-01 and AFP-02) with the antigen was strongly inhibited by calcium-chelating agents. The mAb AFP-11 could be used for estimation of alpha FP by RIA in the concentration range of 5-100 ng/ml. Several combinations of the mAbs could be used in a two-site sandwich-ELISA for similar purposes. Immunoaffinity chromatography on a column of immobilized mAb AFP-01 permitted the purification of alpha FP.