Abstract
The technologies described in this volume enable the isolation of recombi- nant antibodies (Abs) from libraries of variable regions (Fvs) or Fabs displayed on the surface of filamentous phage by fusion to a structural protein (1 -7). Production of selected Abs in sufficient quantity for further evaluation often requires the transfer of their coding sequences to vectors specfiically designed for expression in eukaryotic cells. For Fab and/or immunoglobulin G (IgG) expression, these systems either employ separate plasmids for the heavy-chain (HC) and light-chain (LC) sequences or a single plasmid in which both HC and LC cassettes are expressed.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
oReferences
Barbas, C. F., Kang, A. S., Lerner, R. A., and Benkovic, S. J. (1991) Assembly of combinatorial antibody libraries on phage surfaces: the gene III site. Proc. Natl. Acad. Sci. USA 88, 7978–7982.
Burioni, R., Williamson, R. A., Sanna, P. P., Bloom, F. E., and Burton, D. R. (1994) Recombinant human Fab to glycoprotein D neutralizes infectivity and prevents cell-to-cell transmission of herpes simplex viruses 1 and 2 in vitro. Proc. Natl. Acad. Sci. USA 91, 355–359.
Chanock, R. M., Crowe, J. J., Murphy, B. R., and Burton, D. R. (1993) Human monoclonal antibody Fab fragments cloned from combinatorial libraries: potential usefulness in prevention and/or treatment of major human viral diseases. Infect. Agents Dis. 2, 118–131.
Lerner, R. A., Kang, A. S., Bain, J. D., Burton, D. R., and Barbas, C. F. (1992) Antibodies without immunization. Science 258, 1313–1314.
Sanna, P. P., Williamson, R. A., De Logu, A., Bloom, R. E., and Burton, D. R. (1995) Directed selection of recombinant human monoclonal antibodies to herpes simplex glycoproteins from phage display libraries. Proc. Natl. Acad. Sci. USA 92, 6439–6443.
Williamson, R. A., Burioni, R., Sanna, P. P., Partridge, L. J., Barbas, C. F., and Burton, D. R. (1993) Human monoclonal antibodies against a plethora of viral pathogens from single combinatorial libraries. Proc. Natl. Acad. Sci. USA 90, 4141–4145.
Winter, G., Griffiths, A. D., Hawkins, R. E., and Hoogenboom, H. R. (1994) Making antibodies by phage display technology. Ann. Rev. Immunol. 12, 433–455.
Sanna, P. P., Samson, M. E., Moon, J. S., Rozenshteyn, R., De Logu, A., Williamson, R. A., and Burton, D. R. (1999) pFab-CMV, a single vector system for the rapid conversion of recombinant Fabs into whole IgG1 antibodies. Immunotechnology 4, 185–188.
Sanna, P. P., De Logu, A., Williamson, R. A., Samson, M. E., Altieri, D. C., Bloom, F. E., and Burton, D. R. (1995) Rapid assay of phage-derived recombinant human fabs as bispecific antibodies. Biotechnology 13, 1221–1224.
Harlow, E. and Lane, D. (1988) Antibodies: a Laboratory Manual. Cold Spring Harbor Press, Cold Spring Harbor, NY.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2002 Humana Press Inc.
About this protocol
Cite this protocol
Sanna, P.P. (2002). 35 Expression of Antibody Fab Fragments and Whole Immunoglobulin in Mammalian Cells. In: O’Brien, P.M., Aitken, R. (eds) Antibody Phage Display. Methods in Molecular Biology™, vol 178. Humana Press. https://doi.org/10.1385/1-59259-240-6:389
Download citation
DOI: https://doi.org/10.1385/1-59259-240-6:389
Publisher Name: Humana Press
Print ISBN: 978-0-89603-906-3
Online ISBN: 978-1-59259-240-1
eBook Packages: Springer Protocols