PMID: 6171597Jan 1, 1981Paper

Characterization of a monoclonal antibody to human alpha-fetoprotein and its use in affinity chromatography

Journal of Immunological Methods
U H StenmanJ Schröder

Abstract

A hybrid cell line secreting monoclonal antibodies to human alpha- fetoprotein (AFP) was produced by fusion of a mouse myeloma cell line with spleen cells from a BALB/c mouse immunized with human AFP. The affinity constant of the antibody was about 1.7 X 10(9) l/mol. When clones were grown in vitro, the highest concentration of specific antibody in the culture medium was 25 microgram/ml. A clone was transplanted intraperitoneally into pristine-primed BALB/c recipients. Ascites developed within 3-4 weeks of transplantation, and the maximal antibody concentration in the ascitic fluid was 2.5 mg/ml. The immunoglobulin fraction of ascitic fluid was coupled to cyanogen bromide-activated Sepharose and used for affinity chromatography of AFP. AFP containing less than 1% contaminating proteins was obtained by passing amniotic fluid through the column and eluting the adsorbed AFP with 4 mol/l urea. The monoclonal antibody was used for radioimmunoassay (RIA). The sensitivity obtained was 50 microgram/l, which is adequate for certain clinical applications.

References

Apr 29, 1976·Nature·E Ruoslahti, W D Terry
Sep 30, 1967·Nature·J PorathS Ernback
Dec 28, 1971·Biochimica Et Biophysica Acta·J I Thorell, B G Johansson

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Citations

Jun 24, 1983·Journal of Immunological Methods·M C StuartS R Von Sturmer
May 27, 1983·Journal of Immunological Methods·M HeikinheimoH Bohn
Sep 1, 1983·European Journal of Cancer & Clinical Oncology·B MicheelJ Kopp
Jan 1, 1983·Annals of the New York Academy of Sciences·S Nishi, H Yamazaki
Nov 1, 1984·Scandinavian Journal of Immunology·B NikinmaaJ Schröder
Apr 1, 1983·Bioscience Reports·P HérionA Bollen
Apr 15, 1988·Biochemical and Biophysical Research Communications·E M RutanenT Weber

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