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Automated high throughput microscale antibody purification workflows for accelerating antibody discovery
To rapidly find “best-in-class” antibody therapeutics, it has become essential to develop high throughput (HTP) processes that allow rapid assessment of antibodies for functional and molecular properties. Consequently, it is critical to have access to sufficient amounts of high quality antibody, to...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Taylor & Francis
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5973699/ https://www.ncbi.nlm.nih.gov/pubmed/29494273 http://dx.doi.org/10.1080/19420862.2018.1445450 |
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author | Luan, Peng Lee, Sophia Arena, Tia A. Paluch, Maciej Kansopon, Joe Viajar, Sharon Begum, Zahira Chiang, Nancy Nakamura, Gerald Hass, Philip E. Wong, Athena W. Lazar, Greg A. Gill, Avinash |
author_facet | Luan, Peng Lee, Sophia Arena, Tia A. Paluch, Maciej Kansopon, Joe Viajar, Sharon Begum, Zahira Chiang, Nancy Nakamura, Gerald Hass, Philip E. Wong, Athena W. Lazar, Greg A. Gill, Avinash |
author_sort | Luan, Peng |
collection | PubMed |
description | To rapidly find “best-in-class” antibody therapeutics, it has become essential to develop high throughput (HTP) processes that allow rapid assessment of antibodies for functional and molecular properties. Consequently, it is critical to have access to sufficient amounts of high quality antibody, to carry out accurate and quantitative characterization. We have developed automated workflows using liquid handling systems to conduct affinity-based purification either in batch or tip column mode. Here, we demonstrate the capability to purify >2000 antibodies per day from microscale (1 mL) cultures. Our optimized, automated process for human IgG1 purification using MabSelect SuRe resin achieves ∼70% recovery over a wide range of antibody loads, up to 500 µg. This HTP process works well for hybridoma-derived antibodies that can be purified by MabSelect SuRe resin. For rat IgG2a, which is often encountered in hybridoma cultures and is challenging to purify via an HTP process, we established automated purification with GammaBind Plus resin. Using these HTP purification processes, we can efficiently recover sufficient amounts of antibodies from mammalian transient or hybridoma cultures with quality comparable to conventional column purification. |
format | Online Article Text |
id | pubmed-5973699 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Taylor & Francis |
record_format | MEDLINE/PubMed |
spelling | pubmed-59736992018-05-31 Automated high throughput microscale antibody purification workflows for accelerating antibody discovery Luan, Peng Lee, Sophia Arena, Tia A. Paluch, Maciej Kansopon, Joe Viajar, Sharon Begum, Zahira Chiang, Nancy Nakamura, Gerald Hass, Philip E. Wong, Athena W. Lazar, Greg A. Gill, Avinash MAbs Report To rapidly find “best-in-class” antibody therapeutics, it has become essential to develop high throughput (HTP) processes that allow rapid assessment of antibodies for functional and molecular properties. Consequently, it is critical to have access to sufficient amounts of high quality antibody, to carry out accurate and quantitative characterization. We have developed automated workflows using liquid handling systems to conduct affinity-based purification either in batch or tip column mode. Here, we demonstrate the capability to purify >2000 antibodies per day from microscale (1 mL) cultures. Our optimized, automated process for human IgG1 purification using MabSelect SuRe resin achieves ∼70% recovery over a wide range of antibody loads, up to 500 µg. This HTP process works well for hybridoma-derived antibodies that can be purified by MabSelect SuRe resin. For rat IgG2a, which is often encountered in hybridoma cultures and is challenging to purify via an HTP process, we established automated purification with GammaBind Plus resin. Using these HTP purification processes, we can efficiently recover sufficient amounts of antibodies from mammalian transient or hybridoma cultures with quality comparable to conventional column purification. Taylor & Francis 2018-03-29 /pmc/articles/PMC5973699/ /pubmed/29494273 http://dx.doi.org/10.1080/19420862.2018.1445450 Text en © 2018 The Author(s). Published with license by Taylor & Francis Group, LLC http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives License (http://creativecommons.org/licenses/by-nc-nd/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited, and is not altered, transformed, or built upon in any way. |
spellingShingle | Report Luan, Peng Lee, Sophia Arena, Tia A. Paluch, Maciej Kansopon, Joe Viajar, Sharon Begum, Zahira Chiang, Nancy Nakamura, Gerald Hass, Philip E. Wong, Athena W. Lazar, Greg A. Gill, Avinash Automated high throughput microscale antibody purification workflows for accelerating antibody discovery |
title | Automated high throughput microscale antibody purification workflows for accelerating antibody discovery |
title_full | Automated high throughput microscale antibody purification workflows for accelerating antibody discovery |
title_fullStr | Automated high throughput microscale antibody purification workflows for accelerating antibody discovery |
title_full_unstemmed | Automated high throughput microscale antibody purification workflows for accelerating antibody discovery |
title_short | Automated high throughput microscale antibody purification workflows for accelerating antibody discovery |
title_sort | automated high throughput microscale antibody purification workflows for accelerating antibody discovery |
topic | Report |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5973699/ https://www.ncbi.nlm.nih.gov/pubmed/29494273 http://dx.doi.org/10.1080/19420862.2018.1445450 |
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