Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Lombana, T. Noelle, Dillon, Michael, Bevers III, Jack, Spiess, Christoph
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2015
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
_version_ 1782403977271312384
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
work_keys_str_mv AT lombanatnoelle optimizingantibodyexpressionbyusingthenaturallyoccurringframeworkdiversityinalivebacterialantibodydisplaysystem
AT dillonmichael optimizingantibodyexpressionbyusingthenaturallyoccurringframeworkdiversityinalivebacterialantibodydisplaysystem
AT beversiiijack optimizingantibodyexpressionbyusingthenaturallyoccurringframeworkdiversityinalivebacterialantibodydisplaysystem
AT spiesschristoph optimizingantibodyexpressionbyusingthenaturallyoccurringframeworkdiversityinalivebacterialantibodydisplaysystem