PCR amplification of the functional immunoglobulin heavy chain variable gene from a hybridoma in the presence of two aberrant transcripts

Yazad Irani, Melinda Tea, Ronald Tilton, Douglas J. Coster, Keryn A. Williams, Helen M. Brereton

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Single chain antibody fragment genes are commonly created by splicing together the immunoglobulin light chain (VL) and heavy chain variable (VH) genes of a monoclonal antibody produced by a hybridoma. Selective PCR amplification of the functional immunoglobulin variable gene rearrangements can be complicated by the existence of other unproductive immunoglobulin gene rearrangements in the hybridoma. Here we report the detection and preferential amplification of aberrant transcripts from two unproductive VH gene rearrangements derived from the fusion partner of a hybridoma. The functional VH gene of the monoclonal antibody was successfully amplified by selective use of primers to individual JH segments.

Original languageEnglish (US)
Pages (from-to)246-250
Number of pages5
JournalJournal of Immunological Methods
Volume336
Issue number2
DOIs
StatePublished - Jul 31 2008

Fingerprint

Immunoglobulin Heavy Chain Genes
Gene Rearrangement
Hybridomas
Immunoglobulin Genes
Polymerase Chain Reaction
Monoclonal Antibodies
Genes
Immunoglobulin Light Chains
Single-Chain Antibodies
Immunoglobulin Fragments

Keywords

  • Hybridoma
  • MOPC21
  • Myeloma fusion partner
  • P3X63-Ag8.653
  • scFv
  • Unproductive immunoglobulin rearrangement

ASJC Scopus subject areas

  • Biotechnology
  • Immunology

Cite this

PCR amplification of the functional immunoglobulin heavy chain variable gene from a hybridoma in the presence of two aberrant transcripts. / Irani, Yazad; Tea, Melinda; Tilton, Ronald; Coster, Douglas J.; Williams, Keryn A.; Brereton, Helen M.

In: Journal of Immunological Methods, Vol. 336, No. 2, 31.07.2008, p. 246-250.

Research output: Contribution to journalArticle

Irani, Yazad ; Tea, Melinda ; Tilton, Ronald ; Coster, Douglas J. ; Williams, Keryn A. ; Brereton, Helen M. / PCR amplification of the functional immunoglobulin heavy chain variable gene from a hybridoma in the presence of two aberrant transcripts. In: Journal of Immunological Methods. 2008 ; Vol. 336, No. 2. pp. 246-250.
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