Purification and immunoreactivity of monoclonal antibodies against myeloma-λ chain

Abstract

In this study, two murine IgM-monoclonal antibodies (IgM-McAbs) against λ chain of myeloma protein from hybridoma ascites were purified by Sephadex G200 chromatography. The eluate of the first peak on absorption at 280 nm formed one precipitation band with rabbit anti-mouse IgM or anti-mouse immunoglobulin in agar gel double diffusion test; it also formed a detectable precipitation line in the IgM reaction area in immunoelectrophoresis. These findings showed that the eluate of the first peak on absorption at 280 nm contained purified IgM-McAbs. It was found in indirect ELISA that the immunoreactivity of the eluate of the first peak was four to five fold higher than that of the original ascites with equimolar protein concentration. Double antibody sandwich ELISA analyses of the immunoreactivity of HRP-conjugated IgM-monoclonal antibodies with λ chain denoted that the purified IgM-McAb-HRP-conjugate might be of practical value in quantitative as well as qualitative assay of λ chains.

Keywords

Sephadex G200 chromatography / IgM-McAbs / ELISA

References

[1] PetersJH, et al.. Monoklonale Antikörper, 1985New York, Heidelberg, BerlinSpringer Verlag185-185

[2] WangXL, et al.. Studies on monoclonal anti-isotypic and anti-idiotypic antibodies against leukemia and myeloma: I. Study on monoclonal anti-idiotypic antibodies against chain protein of myeloma. J Tongji Med Univ, 1990, 10(3): 169-169

[3] WeirDM, et al.. Handbook of Experimental Immunology. Immunochemistry, 19884th ed.OxfordBlackwell Scientific Publication13.1-13.1