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Protein Engineering vol. 1 no. 1 pp. 59-65, 1986
© 1986 Oxford University Press

RESEARCH-ARTICLE

The cloning and expression of an anti-peptide antibody: a system for rapid analysis of the binding properties of engineered antibodies

S. Roberts and A.R. Rees

Laboratory of Molecular Biophysics, Department of Zoology, University of Oxford OX1 3QU, UK

The cloning of the light and heavy chain genes of an anti-lysozyme monoclonal antibody is described and the application of site-directed mutagenesis for reconstruction of complete cDNAs from incomplete cDNA clones is discussed. The behaviour of the RNA transcripts produced from these cDNAs after their subcloning into an appropriate Salmonella lyphimurium, SP6, vector and subsequent injection into Xenopus oocytes has been analysed. The antibody secreted by oocytes after RNA injection has been quantitated and its antigen-binding properties compared with wild-type hybridoma antibody. The results show that this route for expression of antibodies, though limited in its capacity to produce more than a few micrograms of protein, is potentially a means for rapid evaluation of the antigen-binding properties of engineered antibodies.

Keywords: anti-peptide antibodies/cDNA cloning/immunoglobulin expression/protein engineering


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