"Inclonals": IgGs and IgG-enzyme fusion proteins produced in an E. coli expression-refolding system.
Full-length antibodies and antibodies that ferry a cargo to target cells are desired biopharmaceuticals. We describe the production of full-length IgGs and IgG-toxin fusion proteins in E. coli. In the presented examples of anti CD30 and anti EGF-receptor antibodies, the antibody heavy and light chains or toxin fusions thereof were expressed in separate bacterial cultures, where they accumulated as insoluble inclusion bodies. Following refolding and purification, high yields (up to 50 mg/L of shake flask culture) of highly purified (>90%) full-length antibodies and antibody-toxin fusions were obtained. The bacterially produced antibodies, named "Inclonals," equaled the performance of the same IgGs that were produced using conventional mammalian cell culture in binding properties as well as in cell killing potency. The rapid and cost effective IgG production process and the high quality of the resultant product may make the bacterial production of full-length IgG and IgG-drug fusion proteins an attractive option for antibody production and a significant contribution to recombinant antibody technology.
Hakim
Rahely
R
Department of Molecular Microbiology and Biotechnology, The George S. Wise Faculty of Life Sciences, Tel Aviv University, Ramat Aviv, Israel.
Benhar
Itai
I
D000818:Animals; D000920:Antibody-Dependent Cell Cytotoxicity; D003362:Cost-Benefit Analysis; D066246:ErbB Receptors / Q000276:immunology; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression / Q000235:genetics / Q000276:immunology; D006801:Humans; D007074:Immunoglobulin G / Q000191:economics / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D000922:Immunotoxins / Q000191:economics / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D002479:Inclusion Bodies / Q000276:immunology / Q000382:microbiology; D017730:Ki-1 Antigen / Q000276:immunology; D011485:Protein Binding; D017510:Protein Folding; D011993:Recombinant Fusion Proteins / Q000191:economics / Q000235:genetics / Q000276:immunology / Q000378:metabolism*
D007074:Immunoglobulin G; D000922:Immunotoxins; D017730:Ki-1 Antigen; D011993:Recombinant Fusion Proteins; C512478:EGFR protein, human; D066246:ErbB Receptors