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Dideoxy mRNA Sequencing as a Method to Analyze Antibody Diversity

  • Gillian M. Griffiths
  • Claudia Berek
  • Matti Kaartinen
  • Cesar Milstein
Part of the Methodological Surveys in Biochemistry and Analysis book series (MSBA, volume 15B)

Abstract

The antigenic specificity of an antibody (Ab) molecule is determined by the amino acid sequence of the light and heavy chain variable regions. We have analyzed the extent to which various possible mechanisms for generating Ab diversity contribute in an immune response to a simple antigen, the hapten 2-phenyloxazolone. In order to identify the relative contributions of the different mechanisms which can generate Ab diversity (e.g. the use of different germ-line V genes, different combinations of V, D and J gene segments and the role of somatic point mutations), nucleotide sequence information was required. We therefore analyzed the V region expression of a series of oxazolone-specific hybridomas by sequencing the mRNA isolated from these cells. The major advantage of this technique is the speed and ease with which the required sequence information can be generated.

Keywords

Combine Site mRNA Template Heavy Chain Variable Region Somatic Point Mutation Nucleotide Sequence Information 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Plenum Press, New York 1985

Authors and Affiliations

  • Gillian M. Griffiths
    • 1
  • Claudia Berek
    • 1
  • Matti Kaartinen
    • 1
  • Cesar Milstein
    • 1
  1. 1.MRC Laboratory of Molecular BiologyCambridgeUK

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