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Optimizing antibody expression by using the naturally occurring framework diversity in a live bacterial antibody display system
Rapid identification of residues that influence antibody expression and thermostability is often needed to move promising therapeutics into the clinic. To establish a method that can assess small expression differences, we developed a Bacterial Antibody Display (BAD) system that overcomes previous l...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4668361/ https://www.ncbi.nlm.nih.gov/pubmed/26631978 http://dx.doi.org/10.1038/srep17488 |
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author | Lombana, T. Noelle Dillon, Michael Bevers III, Jack Spiess, Christoph |
author_facet | Lombana, T. Noelle Dillon, Michael Bevers III, Jack Spiess, Christoph |
author_sort | Lombana, T. Noelle |
collection | PubMed |
description | Rapid identification of residues that influence antibody expression and thermostability is often needed to move promising therapeutics into the clinic. To establish a method that can assess small expression differences, we developed a Bacterial Antibody Display (BAD) system that overcomes previous limitations, enabling the use of full-length formats for antibody and antigen in a live cell setting. We designed a unique library of individual framework variants using natural diversity introduced by somatic hypermutation, and screened half-antibodies for increased expression using BAD. We successfully identify variants that dramatically improve expression yields and in vitro thermostability of two therapeutically relevant antibodies in E. coli and mammalian cells. While we study antibody expression, bacterial display can now be expanded to examine the processes of protein folding and translocation. Additionally, our natural library design strategy could be applied during antibody humanization and library design for in vitro display methods to maintain expression and formulation stability. |
format | Online Article Text |
id | pubmed-4668361 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-46683612015-12-09 Optimizing antibody expression by using the naturally occurring framework diversity in a live bacterial antibody display system Lombana, T. Noelle Dillon, Michael Bevers III, Jack Spiess, Christoph Sci Rep Article Rapid identification of residues that influence antibody expression and thermostability is often needed to move promising therapeutics into the clinic. To establish a method that can assess small expression differences, we developed a Bacterial Antibody Display (BAD) system that overcomes previous limitations, enabling the use of full-length formats for antibody and antigen in a live cell setting. We designed a unique library of individual framework variants using natural diversity introduced by somatic hypermutation, and screened half-antibodies for increased expression using BAD. We successfully identify variants that dramatically improve expression yields and in vitro thermostability of two therapeutically relevant antibodies in E. coli and mammalian cells. While we study antibody expression, bacterial display can now be expanded to examine the processes of protein folding and translocation. Additionally, our natural library design strategy could be applied during antibody humanization and library design for in vitro display methods to maintain expression and formulation stability. Nature Publishing Group 2015-12-03 /pmc/articles/PMC4668361/ /pubmed/26631978 http://dx.doi.org/10.1038/srep17488 Text en Copyright © 2015, Macmillan Publishers Limited http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ |
spellingShingle | Article Lombana, T. Noelle Dillon, Michael Bevers III, Jack Spiess, Christoph Optimizing antibody expression by using the naturally occurring framework diversity in a live bacterial antibody display system |
title | Optimizing antibody expression by using the naturally occurring framework diversity in a live bacterial antibody display system |
title_full | Optimizing antibody expression by using the naturally occurring framework diversity in a live bacterial antibody display system |
title_fullStr | Optimizing antibody expression by using the naturally occurring framework diversity in a live bacterial antibody display system |
title_full_unstemmed | Optimizing antibody expression by using the naturally occurring framework diversity in a live bacterial antibody display system |
title_short | Optimizing antibody expression by using the naturally occurring framework diversity in a live bacterial antibody display system |
title_sort | optimizing antibody expression by using the naturally occurring framework diversity in a live bacterial antibody display system |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4668361/ https://www.ncbi.nlm.nih.gov/pubmed/26631978 http://dx.doi.org/10.1038/srep17488 |
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