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

Here we describe a facile and robust genetic selection for isolating full-length IgG antibodies from combinatorial libraries expressed in the cytoplasm of redox-engineered Escherichia coli cells. The method is based on the transport of a bifunctional substrate comprised of an antigen fused to chlora...

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Autores principales: Robinson, Michael-Paul, Jung, Jinjoo, Lopez-Barbosa, Natalia, Chang, Matthew, Li, Mingji, Jaroentomeechai, Thapakorn, Cox, Emily C., Zheng, Xiaolu, Berkmen, Mehmet, DeLisa, Matthew P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10267130/
https://www.ncbi.nlm.nih.gov/pubmed/37316535
http://dx.doi.org/10.1038/s41467-023-39178-x
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author Robinson, Michael-Paul
Jung, Jinjoo
Lopez-Barbosa, Natalia
Chang, Matthew
Li, Mingji
Jaroentomeechai, Thapakorn
Cox, Emily C.
Zheng, Xiaolu
Berkmen, Mehmet
DeLisa, Matthew P.
author_facet Robinson, Michael-Paul
Jung, Jinjoo
Lopez-Barbosa, Natalia
Chang, Matthew
Li, Mingji
Jaroentomeechai, Thapakorn
Cox, Emily C.
Zheng, Xiaolu
Berkmen, Mehmet
DeLisa, Matthew P.
author_sort Robinson, Michael-Paul
collection PubMed
description Here we describe a facile and robust genetic selection for isolating full-length IgG antibodies from combinatorial libraries expressed in the cytoplasm of redox-engineered Escherichia coli cells. 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 utility of this approach is first demonstrated by isolating affinity-matured cyclonal variants that specifically bind their cognate antigen, the leucine zipper domain of a yeast transcriptional activator, with subnanomolar affinities, which represent a ~20-fold improvement over the parental IgG. We then use the genetic assay to discover antigen-specific cyclonals from a naïve human antibody repertoire, leading to the identification of lead IgG candidates with affinity and specificity for an influenza hemagglutinin-derived peptide antigen.
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spelling pubmed-102671302023-06-15 Isolation of full-length IgG antibodies from combinatorial libraries expressed in the cytoplasm of Escherichia coli Robinson, Michael-Paul Jung, Jinjoo Lopez-Barbosa, Natalia Chang, Matthew Li, Mingji Jaroentomeechai, Thapakorn Cox, Emily C. Zheng, Xiaolu Berkmen, Mehmet DeLisa, Matthew P. Nat Commun Article Here we describe a facile and robust genetic selection for isolating full-length IgG antibodies from combinatorial libraries expressed in the cytoplasm of redox-engineered Escherichia coli cells. 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 utility of this approach is first demonstrated by isolating affinity-matured cyclonal variants that specifically bind their cognate antigen, the leucine zipper domain of a yeast transcriptional activator, with subnanomolar affinities, which represent a ~20-fold improvement over the parental IgG. We then use the genetic assay to discover antigen-specific cyclonals from a naïve human antibody repertoire, leading to the identification of lead IgG candidates with affinity and specificity for an influenza hemagglutinin-derived peptide antigen. Nature Publishing Group UK 2023-06-14 /pmc/articles/PMC10267130/ /pubmed/37316535 http://dx.doi.org/10.1038/s41467-023-39178-x Text en © This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Robinson, Michael-Paul
Jung, Jinjoo
Lopez-Barbosa, Natalia
Chang, Matthew
Li, Mingji
Jaroentomeechai, Thapakorn
Cox, Emily C.
Zheng, Xiaolu
Berkmen, Mehmet
DeLisa, Matthew P.
Isolation of full-length IgG antibodies from combinatorial libraries expressed in the cytoplasm of Escherichia coli
title Isolation of full-length IgG antibodies from combinatorial libraries expressed in the cytoplasm of Escherichia coli
title_full Isolation of full-length IgG antibodies from combinatorial libraries expressed in the cytoplasm of Escherichia coli
title_fullStr Isolation of full-length IgG antibodies from combinatorial libraries expressed in the cytoplasm of Escherichia coli
title_full_unstemmed Isolation of full-length IgG antibodies from combinatorial libraries expressed in the cytoplasm of Escherichia coli
title_short Isolation of full-length IgG antibodies from combinatorial libraries expressed in the cytoplasm of Escherichia coli
title_sort isolation of full-length igg antibodies from combinatorial libraries expressed in the cytoplasm of escherichia coli
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10267130/
https://www.ncbi.nlm.nih.gov/pubmed/37316535
http://dx.doi.org/10.1038/s41467-023-39178-x
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