Recombinant human antibodies: linkage of an Fab fragment from a combinatorial library to an Fc fragment for expression in mammalian cell culture

Article type: Research Article

Authors: Bender, Eckhard^{a; b} | Woof, Jennifer M.^a | Atkin, Julie D.^a | Barker, Mike D.^a | Bebbington, Chris R.^c | Burton, Dennis R.^{a; b;}

Affiliations: [a] Krebs Institute, Department of Molecular Biology and Biotechnology, University of Sheffield, Sheffield, UK | [b] Departments of Immunology and Molecular Biology, Scripps Research Institute, La Jolla, CA, USA | [c] Celltech Ltd., Slough, UK

Note: [] Address reprint requests to Dr. Dennis R. Burton, The Scripps Research Institute, 10666 North Torrey Pines Road, La Jolla, CA 92037.

Abstract: The combinatorial phage library approach to immunoglobulin repertoire cloning recently made it possible to isolate gene fragments encoding human immunoglobulin G1 Fabs binding with high affinity to specific antigens. Here we describe the construction of genes encoding whole human anti–tetanus toxoid antibodies based on one of these gene fragments and the efficient expression of these constructs by co-transfection of separate heavy and light chain vectors into a Chinese hamster ovary cell line constitutively expressing a viral transactivator protein. This system will be generally useful for the rapid analysis of recombinant antibodies derived from combinatorial libraries.

Keywords: human antibody, immunoglobulin repertoire cloning, CHO cells, tetanus toxoid

DOI: 10.3233/HAB-1993-4205

Journal: Human Antibodies, vol. 4, no. 2, pp. 74-79, 1993