Abstract
A plasmid foroptimizedproteinexpression of recombinant Fv antibodies (pOPE) inE. coli was used to express the variable domains of the murine monoclonal antibody HD39 specific for the human B-cell surface antigen CD22. The production of Fv antibodies by pOPE can be regulated over a wide range by varying the IPTG concentration. Antibodies that can discriminate between secreted and nonsecreted Fv antibody fragments were used to show that secretion is the limiting step for the production of functional Fv antibodies. IPTG concentrations above 20 μM increased the total antibody production, but did not yield larger amounts of secreted Fv antibodies. The addition of five histidines to the C terminus facilitates an easy single-step enrichment procedure based on immobilized metal affinity chromatography.
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Dübel, S., Breitling, F., Klewinghaus, I. et al. Regulated secretion and purification of recombinant antibodies inE. coli . Cell Biophysics 21, 69–79 (1992). https://doi.org/10.1007/BF02789479
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DOI: https://doi.org/10.1007/BF02789479