Multiplex screening for functionally rearranged immunoglobulin variable regions reveals expression of hybridoma-specific aberrant V-genes

J Immunol Methods. 2001 Mar 1;249(1-2):245-52. doi: 10.1016/s0022-1759(00)00337-9.

Abstract

Modification of antibody effector functions is commonly performed by chimerization or humanization. Cloning of antibody variable regions from hybridomas represents a first step that is frequently hampered by the expression of non-functionally rearranged variable regions in hybridoma cells that originate from MOPC21-derived fusion partners. We now present a simple method to clone functionally rearranged V-genes, based on V-gene-specific multiplex PCR screening. Using this method we document the expression of aberrant V-genes that originate from the original B-cell used for the hybridoma generation, not from the fusion partner, and are - thus - hybridoma specific.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Gene Rearrangement, B-Lymphocyte
  • Genes, Immunoglobulin*
  • Hybridomas
  • Immunoglobulin Variable Region / genetics*
  • Immunologic Techniques*
  • Mice
  • Molecular Sequence Data

Substances

  • Immunoglobulin Variable Region