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Carl Borrebaeck

Carl Borrebaeck

Professor

Carl Borrebaeck

Rapid cloning of rearranged immunoglobulin genes from human hybridoma cells using mixed primers and the polymerase chain reaction

Author

  • James W Larrick
  • Lena Danielsson
  • Carol A Brenner
  • Magnus Abrahamson
  • Kirk E Fry
  • Carl Borrebaeck

Summary, in English

A general method to directly obtain the DNA sequence of the variable regions of any immunoglobulin chain using a mixture of oligomer primers and the polymerase chain reaction (PCR) is described. Mixed oligonucleotide primers corresponding to the 5′ signal peptide and a conserved 3′ constant region primer were used for enzymatic amplification of each of the heavy and light chain variable regions of a human hybridoma producing a monoclonal antibody recognizing an epitope of gp120 of the human immunodeficiency virus 1. The amplified DNA segments were cloned and the sequence was determined for the heavy chain variable region. This method will greatly facilitate structural and functional studies of immunoglobulins by reducing the effort to clone and sequence the members of the immunoglobulin as well as other multigene families.

Department/s

  • Division of Clinical Chemistry and Pharmacology
  • Department of Immunotechnology

Publishing year

1989

Language

English

Pages

1250-1256

Publication/Series

Biochemical and Biophysical Research Communications

Volume

160

Issue

3

Document type

Journal article

Publisher

Elsevier

Topic

  • Biological Sciences

Status

Published

ISBN/ISSN/Other

  • ISSN: 1090-2104