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).
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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