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Isolation of full-length IgG antibodies from combinatorial libraries expressed in the cytoplasm of Escherichia coli

Michael-Paul Robinson, Emily C. Cox, Mingji Li, View ORCID ProfileThapakorn Jaroentomeechai, Xiaolu Zheng, Matthew Chang, View ORCID ProfileMehmet Berkmen, View ORCID ProfileMatthew P. DeLisa
doi: https://doi.org/10.1101/2020.05.09.085944
Michael-Paul Robinson
1Robert F. Smith School of Chemical and Biomolecular Engineering, Cornell University, Ithaca, NY 14853 USA
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Emily C. Cox
1Robert F. Smith School of Chemical and Biomolecular Engineering, Cornell University, Ithaca, NY 14853 USA
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Mingji Li
1Robert F. Smith School of Chemical and Biomolecular Engineering, Cornell University, Ithaca, NY 14853 USA
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Thapakorn Jaroentomeechai
1Robert F. Smith School of Chemical and Biomolecular Engineering, Cornell University, Ithaca, NY 14853 USA
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  • ORCID record for Thapakorn Jaroentomeechai
Xiaolu Zheng
1Robert F. Smith School of Chemical and Biomolecular Engineering, Cornell University, Ithaca, NY 14853 USA
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Matthew Chang
1Robert F. Smith School of Chemical and Biomolecular Engineering, Cornell University, Ithaca, NY 14853 USA
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Mehmet Berkmen
2New England Biolabs, 240 County Road, Ipswich, Massachusetts 01938, USA
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  • ORCID record for Mehmet Berkmen
Matthew P. DeLisa
1Robert F. Smith School of Chemical and Biomolecular Engineering, Cornell University, Ithaca, NY 14853 USA
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  • ORCID record for Matthew P. DeLisa
  • For correspondence: md255@cornell.edu
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Abstract

We describe a facile and robust genetic selection for isolating full-length IgG antibodies from combinatorial libraries expressed in the cytoplasm of the genetically engineered Escherichia coli strain, SHuffle. The method is based on the transport of a bifunctional substrate comprised of an antigen fused to chloramphenicol acetyltransferase, which allows positive selection of bacterial cells co-expressing cytoplasmic IgGs called ‘cyclonals’ that specifically capture the chimeric antigen and sequester the antibiotic resistance marker in the cytoplasm. The selective power of this approach was demonstrated by facile isolation of novel complementarity-determining regions for a cyclonal that specifically recognized the basic-region leucine zipper domain of the yeast transcriptional activator protein Gcn4.

Competing Interest Statement

M.P.D. has ownership interest (including stock, patents, etc.) in SwiftScale Biologics, Inc. M.P.D.’s interests are reviewed and managed by Cornell University in accordance with their conflict of interest policies. M.B. is employed by NEB, which commercializes SHuffle cells. All other authors declare no competing interests.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted May 10, 2020.
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Isolation of full-length IgG antibodies from combinatorial libraries expressed in the cytoplasm of Escherichia coli
Michael-Paul Robinson, Emily C. Cox, Mingji Li, Thapakorn Jaroentomeechai, Xiaolu Zheng, Matthew Chang, Mehmet Berkmen, Matthew P. DeLisa
bioRxiv 2020.05.09.085944; doi: https://doi.org/10.1101/2020.05.09.085944
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Isolation of full-length IgG antibodies from combinatorial libraries expressed in the cytoplasm of Escherichia coli
Michael-Paul Robinson, Emily C. Cox, Mingji Li, Thapakorn Jaroentomeechai, Xiaolu Zheng, Matthew Chang, Mehmet Berkmen, Matthew P. DeLisa
bioRxiv 2020.05.09.085944; doi: https://doi.org/10.1101/2020.05.09.085944

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