Efficient expression of full-length antibodies in the cytoplasm of engineered bacteria

Michael-Paul Robinson, Na Ke, Julie Lobstein, Cristen Peterson, Alana Szkodny, Thomas J. Mansell, Corinna Tuckey, Paul D. Riggs, Paul A. Colussi, Christopher J. Noren, Christopher H. Taron, Matthew P. DeLisa () and Mehmet Berkmen ()
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Michael-Paul Robinson: School of Chemical and Biomolecular Engineering, Cornell University
Na Ke: New England Biolabs
Julie Lobstein: New England Biolabs
Cristen Peterson: School of Chemical and Biomolecular Engineering, Cornell University
Alana Szkodny: School of Chemical and Biomolecular Engineering, Cornell University
Thomas J. Mansell: School of Chemical and Biomolecular Engineering, Cornell University
Corinna Tuckey: New England Biolabs
Paul D. Riggs: New England Biolabs
Paul A. Colussi: New England Biolabs
Christopher J. Noren: New England Biolabs
Christopher H. Taron: New England Biolabs
Matthew P. DeLisa: School of Chemical and Biomolecular Engineering, Cornell University
Mehmet Berkmen: New England Biolabs

Nature Communications, 2015, vol. 6, issue 1, 1-9

Abstract: Abstract Current methods for producing immunoglobulin G (IgG) antibodies in engineered cells often require refolding steps or secretion across one or more biological membranes. Here, we describe a robust expression platform for biosynthesis of full-length IgG antibodies in the Escherichia coli cytoplasm. Synthetic heavy and light chains, both lacking canonical export signals, are expressed in specially engineered E. coli strains that permit formation of stable disulfide bonds within the cytoplasm. IgGs with clinically relevant antigen- and effector-binding activities are readily produced in the E. coli cytoplasm by grafting antigen-specific variable heavy and light domains into a cytoplasmically stable framework and remodelling the fragment crystallizable domain with amino-acid substitutions that promote binding to Fcγ receptors. The resulting cytoplasmic IgGs—named ‘cyclonals’—effectively bypass the potentially rate-limiting steps of membrane translocation and glycosylation.

Date: 2015
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DOI: 10.1038/ncomms9072

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