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Journal of Tongji Medical University

, Volume 11, Issue 4, pp 204–207 | Cite as

Purification and immunoreactivity of monoclonal antibodies against myeloma-λ chain

  • Shen Guan-xin
  • Zhu Hui-fen
  • Su Na
  • Wang Xiao-lin
  • Zhang Yue
  • Yang Dao-feng
Article
  • 14 Downloads

Summary

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.

Key words

Sephadex G200 chromatography IgM-McAbs ELISA 

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References

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    Peters JH, et al. Monoklonale Antikörper. New York, Heidelberg, Berlin: Springer Verlag, 1985:185Google Scholar
  2. 2.
    Wang XL, 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): 169PubMedGoogle Scholar
  3. 3.
    Weir DM, et al. Handbook of Experimental Immunology. 4th ed. Volume 1. Immunochemistry. Oxford: Blackwell Scientific Publication, 1988:13.1Google Scholar

Copyright information

© Springer 1991

Authors and Affiliations

  • Shen Guan-xin
    • 1
  • Zhu Hui-fen
    • 1
  • Su Na
    • 1
  • Wang Xiao-lin
    • 1
  • Zhang Yue
    • 1
  • Yang Dao-feng
    • 2
  1. 1.Research Center of Experimental MedicineTongji Medical UniversityWuhan
  2. 2.Department of Infectious Diseases, Tongji HospitalTongji Medical UniversityWuhan

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