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Understanding ForteBio’s Sensors for High-Throughput Kinetic and Epitope Screening for Purified Antibodies and Yeast Culture Supernatant
Real-time and label-free antibody screening systems are becoming more popular because of the increasing output of purified antibodies and antibody supernatant from many antibody discovery platforms. However, the properties of the biosensor can greatly affect the kinetic and epitope binning results g...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
SAGE Publications
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4708621/ https://www.ncbi.nlm.nih.gov/pubmed/26442912 http://dx.doi.org/10.1177/1087057115609564 |
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author | Yu, Yao Mitchell, Scott Lynaugh, Heather Brown, Michael Nobrega, R. Paul Zhi, Xiaoyong Sun, Tingwan Caffry, Isabelle Cao, Yuan Yang, Rong Burnina, Irina Xu, Yingda Estep, Patricia |
author_facet | Yu, Yao Mitchell, Scott Lynaugh, Heather Brown, Michael Nobrega, R. Paul Zhi, Xiaoyong Sun, Tingwan Caffry, Isabelle Cao, Yuan Yang, Rong Burnina, Irina Xu, Yingda Estep, Patricia |
author_sort | Yu, Yao |
collection | PubMed |
description | Real-time and label-free antibody screening systems are becoming more popular because of the increasing output of purified antibodies and antibody supernatant from many antibody discovery platforms. However, the properties of the biosensor can greatly affect the kinetic and epitope binning results generated by these label-free screening systems. ForteBio human-specific ProA, anti-human IgG quantitation (AHQ), anti-human Fc capture (AHC) sensors, and custom biotinylated-anti-human Fc capture (b-AHFc) sensors were evaluated in terms of loading ability, regeneration, kinetic characterization, and epitope binning with both purified IgG and IgG supernatant. AHC sensors proved unreliable for kinetic or binning assays at times, whereas AHQ sensors showed poor loading and regeneration abilities. ProA sensors worked well with both purified IgG and IgG supernatant. However, the interaction between ProA sensors and the Fab region of the IgG with VH3 germline limited the application of ProA sensors, especially in the epitope binning experiment. In an attempt to generate a biosensor type that would be compatible with a variety of germlines and sample types, we found that the custom b-AHFc sensors appeared to be robust working with both purified IgG and IgG supernatant, with little evidence of sensor-related artifacts. |
format | Online Article Text |
id | pubmed-4708621 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | SAGE Publications |
record_format | MEDLINE/PubMed |
spelling | pubmed-47086212016-01-20 Understanding ForteBio’s Sensors for High-Throughput Kinetic and Epitope Screening for Purified Antibodies and Yeast Culture Supernatant Yu, Yao Mitchell, Scott Lynaugh, Heather Brown, Michael Nobrega, R. Paul Zhi, Xiaoyong Sun, Tingwan Caffry, Isabelle Cao, Yuan Yang, Rong Burnina, Irina Xu, Yingda Estep, Patricia J Biomol Screen Technical Notes Real-time and label-free antibody screening systems are becoming more popular because of the increasing output of purified antibodies and antibody supernatant from many antibody discovery platforms. However, the properties of the biosensor can greatly affect the kinetic and epitope binning results generated by these label-free screening systems. ForteBio human-specific ProA, anti-human IgG quantitation (AHQ), anti-human Fc capture (AHC) sensors, and custom biotinylated-anti-human Fc capture (b-AHFc) sensors were evaluated in terms of loading ability, regeneration, kinetic characterization, and epitope binning with both purified IgG and IgG supernatant. AHC sensors proved unreliable for kinetic or binning assays at times, whereas AHQ sensors showed poor loading and regeneration abilities. ProA sensors worked well with both purified IgG and IgG supernatant. However, the interaction between ProA sensors and the Fab region of the IgG with VH3 germline limited the application of ProA sensors, especially in the epitope binning experiment. In an attempt to generate a biosensor type that would be compatible with a variety of germlines and sample types, we found that the custom b-AHFc sensors appeared to be robust working with both purified IgG and IgG supernatant, with little evidence of sensor-related artifacts. SAGE Publications 2016-01 /pmc/articles/PMC4708621/ /pubmed/26442912 http://dx.doi.org/10.1177/1087057115609564 Text en © 2015 Society for Laboratory Automation and Screening http://creativecommons.org/licenses/by-nc/3.0/ This article is distributed under the terms of the Creative Commons Attribution-NonCommercial 3.0 License (http://www.creativecommons.org/licenses/by-nc/3.0/) which permits non-commercial use, reproduction and distribution of the work without further permission provided the original work is attributed as specified on the SAGE and Open Access page(https://us.sagepub.com/en-us/nam/open-access-at-sage). |
spellingShingle | Technical Notes Yu, Yao Mitchell, Scott Lynaugh, Heather Brown, Michael Nobrega, R. Paul Zhi, Xiaoyong Sun, Tingwan Caffry, Isabelle Cao, Yuan Yang, Rong Burnina, Irina Xu, Yingda Estep, Patricia Understanding ForteBio’s Sensors for High-Throughput Kinetic and Epitope Screening for Purified Antibodies and Yeast Culture Supernatant |
title | Understanding ForteBio’s Sensors for High-Throughput Kinetic and Epitope Screening for Purified Antibodies and Yeast Culture Supernatant |
title_full | Understanding ForteBio’s Sensors for High-Throughput Kinetic and Epitope Screening for Purified Antibodies and Yeast Culture Supernatant |
title_fullStr | Understanding ForteBio’s Sensors for High-Throughput Kinetic and Epitope Screening for Purified Antibodies and Yeast Culture Supernatant |
title_full_unstemmed | Understanding ForteBio’s Sensors for High-Throughput Kinetic and Epitope Screening for Purified Antibodies and Yeast Culture Supernatant |
title_short | Understanding ForteBio’s Sensors for High-Throughput Kinetic and Epitope Screening for Purified Antibodies and Yeast Culture Supernatant |
title_sort | understanding fortebio’s sensors for high-throughput kinetic and epitope screening for purified antibodies and yeast culture supernatant |
topic | Technical Notes |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4708621/ https://www.ncbi.nlm.nih.gov/pubmed/26442912 http://dx.doi.org/10.1177/1087057115609564 |
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