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Antibody engineering using Escherichia coli as host

Antibody engineering using Escherichia coli as host
Antibody engineering using Escherichia coli as host

This chapter discusses the expression and genetic manipulation of antibody fragments in E. coli. The use of the polymerase chain reaction (PCR) for the isolation of antibody variable domain genes is described. The expression of immunoglobulin fragments with antigen binding activities in E. coli is now routinely possible. Using such expression systems, Fv, Fab, and scFv fragments and single VH domains can be produced as secreted proteins in yields of the order of milligrams per liter. Expression systems are developed for the production of antibody scFv or Fab fragments by repertoire cloning followed by selection. Diverse repertoires of genes encoding variable domains (VH and VL) can be isolated by the PCR and cloned for expression using these systems, which allow the selection of recombinants producing fragments with the desired antigen-binding specificities. This technology is rapidly evolving, and coupled with the development of systems for the random rnutagenesis and selection of higher-affinity antibody fragments, could provide an alternative rapid route to hybridoma technology.

1054-3589
1-20
Ward, E. Sally
b31c0877-8abe-485f-b800-244a9d3cd6cc
Ward, E. Sally
b31c0877-8abe-485f-b800-244a9d3cd6cc

Ward, E. Sally (1993) Antibody engineering using Escherichia coli as host. Advances in Pharmacology, 24 (C), 1-20. (doi:10.1016/S1054-3589(08)60931-X).

Record type: Article

Abstract

This chapter discusses the expression and genetic manipulation of antibody fragments in E. coli. The use of the polymerase chain reaction (PCR) for the isolation of antibody variable domain genes is described. The expression of immunoglobulin fragments with antigen binding activities in E. coli is now routinely possible. Using such expression systems, Fv, Fab, and scFv fragments and single VH domains can be produced as secreted proteins in yields of the order of milligrams per liter. Expression systems are developed for the production of antibody scFv or Fab fragments by repertoire cloning followed by selection. Diverse repertoires of genes encoding variable domains (VH and VL) can be isolated by the PCR and cloned for expression using these systems, which allow the selection of recombinants producing fragments with the desired antigen-binding specificities. This technology is rapidly evolving, and coupled with the development of systems for the random rnutagenesis and selection of higher-affinity antibody fragments, could provide an alternative rapid route to hybridoma technology.

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Published date: 1 January 1993

Identifiers

Local EPrints ID: 425262
URI: https://eprints.soton.ac.uk/id/eprint/425262
ISSN: 1054-3589
PURE UUID: 5915061f-23fa-4ab8-a6f6-c2e71e26ee95
ORCID for E. Sally Ward: ORCID iD orcid.org/0000-0003-3232-7238

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Date deposited: 12 Oct 2018 16:30
Last modified: 14 Mar 2019 01:21

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Author: E. Sally Ward ORCID iD

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