PMID: 7580911Apr 1, 1995Paper

Specific amplification by PCR of rearranged genomic variable regions of immunoglobulin genes from mouse hybridoma cells

PCR Methods and Applications
J BerdozJ P Kraehenbuhl

Abstract

We have designed a novel strategy for the isolation of the rearranged genomic fragments encoding the L-VH-D-JH and L-V kappa/lambda-J kappa/lambda regions of mouse immunoglobulin genes. This strategy is based on the PCR amplification of genomic DNA from mouse hybridomas using multiple specific primers chosen in the 5'-untranslated region and in the intron downstream of the rearranged JH/J kappa/lambda sequences. Variable regions with intact coding sequences, including full-length leader peptides (L) can be obtained without previous DNA sequencing. Our strategy is based on a genomic template that produces fragments that do not need to be adapted for recombinant antibody expression, thus facilitating the generation of chimeric and isotype-switched immunoglobulins.

Citations

Jan 30, 2002·Trends in Biotechnology·Blaise Corthésy
Mar 17, 1999·Proceedings of the National Academy of Sciences of the United States of America·J BerdozB Corthésy
Dec 30, 2014·Monoclonal Antibodies in Immunodiagnosis and Immunotherapy·Magdalena BialonChristiane Püttmann
Jun 1, 2018·SLAS Discovery·Zhaoping Liu, John O'Rourke
Dec 30, 1999·Biological Chemistry·B Corthésy, F Spertini
Mar 6, 2008·Applied Microbiology and Biotechnology·Simon KorenVladka Curin Serbec

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