Molecular characterization of hybridoma subclones spontaneously switching at high frequencies in vitro

Maria D. Iglesias-Ussel, Jiri Zavadil, Matthew D. Scharff

Research output: Contribution to journalArticle

2 Scopus citations

Abstract

The hybridoma technology allows the production of large quantities of specific antibodies of a single isotype. Since different isotypes have special effector functions and are distributed distinctively throughout the body, it is often useful to have a library of switch variants from the original monoclonal antibody. We have shown previously that forced expression of activation induced cytidine deaminase (AID) in hybridomas increased their very low frequency of class switch recombination (CSR) in vitro only ∼ 7-13 fold. Since we had previously identified rare hybridoma subclones that spontaneously switched at more than 100 times higher frequencies, we have now examined those higher switching variants to search for ways to further increase the frequency of isotype switching in vitro. AID was not responsible for the ∼ 100 fold increase in CSR, so we used whole-genome gene expression profiling to provide a platform for studying candidate molecular pathways underlying spontaneous CSR in hybridomas.

Original languageEnglish (US)
Pages (from-to)71-78
Number of pages8
JournalJournal of Immunological Methods
Volume350
Issue number1-2
DOIs
StatePublished - Oct 31 2009

Keywords

  • AID
  • Class switch recombination
  • Hybridoma
  • Immunoglobulin
  • Microarrays
  • Real-time RT-PCR

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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