Antibody engineering using Escherichia coli as host.
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. Moreover, expression systems are being rapidly developed for the production of antibody scFv or Fab fragments by repertoire cloning followed by selection. Diverse repertoires of genes encoding VH and VL domains can be isolated by the PCR and cloned for expression using these systems, which allow the selection of recombinants that produce fragments with the desired antigen binding specificities. This technology is rapidly evolving and, coupled with the development of systems for the random mutagenesis and selection of higher-affinity antibody fragments, could, in the longer term, provide an alternative rapid route to hybridoma technology.
Cancer Immunobiology Center, University of Texas Southwestern Medical Center, Dallas 75235.
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Ward
D000818:Animals; D000906:Antibodies / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics* / Q000276:immunology*; D005818:Genetic Engineering; D006801:Humans
D000906:Antibodies