Increased frequency of both total and specific monoclonal antibody producing hybridomas using a fusion partner that constitutively expresses recombinant IL-6

J Immunol Methods. 1992 Apr 8;148(1-2):199-207. doi: 10.1016/0022-1759(92)90173-q.

Abstract

The addition of auxiliary feeder cells or conditioned medium has been shown to augment the yield of mouse hybridomas obtained following the cell-cell fusion of myeloma and B lymphocytes. The addition of one of these factors, interleukin-6 (IL-6) has been found to increase the proportion of hybridomas secreting monoclonal antibodies of desired specificity. As an alternative genetic approach, we have examined the efficacy of a retroviral infectant of Sp2/0 cells that constitutively expresses recombinant murine IL-6 (Sp2/mIL-6) as fusion partner. The results demonstrated that the yields of both viable Ig-secreting hybridomas, and antigen-specific monoclonal antibodies were increased 3-15-fold and 5-9-fold, respectively, with the Sp2/mIL-6 relative to Sp2/0 or Sp2/neo cells as fusion partner. Sp2/mIL-6 cells generated hybridomas with comparable growth rates, stability, and Ig production. The results of staining nascent hybridoma colonies immunohistochemically for Ig production suggest that Sp2/mIL-6 cells as a fusion partner increased the viability and/or stability of nascent hybrid cells that are producing Ig. Thus the Sp2/mIL-6 cells are an improved myeloma parent for the generation of large numbers of antibody-producing hybridomas against specific antigens.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal / biosynthesis
  • B-Lymphocytes / immunology
  • Blotting, Northern
  • Blotting, Southern
  • Cell Fusion
  • Chromosome Mapping
  • DNA / analysis
  • Genetic Vectors
  • Humans
  • Hybridomas / immunology*
  • Immunoglobulins / analysis
  • Interleukin-6 / pharmacology*
  • Mice
  • Mice, Inbred BALB C
  • Multiple Myeloma / immunology
  • RNA / analysis
  • Recombinant Proteins / pharmacology
  • Retroviridae

Substances

  • Antibodies, Monoclonal
  • Immunoglobulins
  • Interleukin-6
  • Recombinant Proteins
  • RNA
  • DNA