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Bispecific antibody target pair discovery by high-throughput phenotypic screening using in vitro combinatorial Fab libraries

Bispecific antibodies can uniquely influence cellular responses, but selecting target combinations for optimal functional activity remains challenging. Here we describe a high-throughput, combinatorial, phenotypic screening approach using a new bispecific antibody target discovery format, allowing s...

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Autores principales: Bhatta, Pallavi, Whale, Kevin D., Sawtell, Amy K., Thompson, Clare L., Rapecki, Stephen E., Cook, David A., Twomey, Breda M., Mennecozzi, Milena, Starkie, Laura E., Barry, Emily M. C., Peters, Shirley J., Kamal, Ahmad M., Finney, Helene M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Taylor & Francis 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7849716/
https://www.ncbi.nlm.nih.gov/pubmed/33487120
http://dx.doi.org/10.1080/19420862.2020.1859049
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author Bhatta, Pallavi
Whale, Kevin D.
Sawtell, Amy K.
Thompson, Clare L.
Rapecki, Stephen E.
Cook, David A.
Twomey, Breda M.
Mennecozzi, Milena
Starkie, Laura E.
Barry, Emily M. C.
Peters, Shirley J.
Kamal, Ahmad M.
Finney, Helene M.
author_facet Bhatta, Pallavi
Whale, Kevin D.
Sawtell, Amy K.
Thompson, Clare L.
Rapecki, Stephen E.
Cook, David A.
Twomey, Breda M.
Mennecozzi, Milena
Starkie, Laura E.
Barry, Emily M. C.
Peters, Shirley J.
Kamal, Ahmad M.
Finney, Helene M.
author_sort Bhatta, Pallavi
collection PubMed
description Bispecific antibodies can uniquely influence cellular responses, but selecting target combinations for optimal functional activity remains challenging. Here we describe a high-throughput, combinatorial, phenotypic screening approach using a new bispecific antibody target discovery format, allowing screening of hundreds of target combinations. Simple in vitro mixing of Fab-fusion proteins from a diverse library enables the generation of thousands of screen-ready bispecific antibodies for high-throughput, biologically relevant assays. We identified an obligate bispecific co-targeting CD79a/b and CD22 as a potent inhibitor of human B cell activation from a short-term flow cytometry signaling assay. A long-term, high-content imaging assay identified anti-integrin bispecific inhibitors of human cell matrix accumulation targeting integrins β1 and β6 or αV and β1. In all cases, functional activity was conserved from the bispecific screening format to a therapeutically relevant format. We also introduce a broader type of mechanistic screen whereby functional modulation of different cell subsets in peripheral blood mononuclear cells was evaluated simultaneously. We identified bispecific antibodies capable of activating different T cell subsets of potential interest for applications in oncology or infectious disease, as well as bispecifics abrogating T cell activity of potential interest to autoimmune or inflammatory disease. The bispecific target pair discovery technology described herein offers access to new target biology and unique bispecific therapeutic opportunities in diverse disease indications.
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spelling pubmed-78497162021-02-05 Bispecific antibody target pair discovery by high-throughput phenotypic screening using in vitro combinatorial Fab libraries Bhatta, Pallavi Whale, Kevin D. Sawtell, Amy K. Thompson, Clare L. Rapecki, Stephen E. Cook, David A. Twomey, Breda M. Mennecozzi, Milena Starkie, Laura E. Barry, Emily M. C. Peters, Shirley J. Kamal, Ahmad M. Finney, Helene M. MAbs Report Bispecific antibodies can uniquely influence cellular responses, but selecting target combinations for optimal functional activity remains challenging. Here we describe a high-throughput, combinatorial, phenotypic screening approach using a new bispecific antibody target discovery format, allowing screening of hundreds of target combinations. Simple in vitro mixing of Fab-fusion proteins from a diverse library enables the generation of thousands of screen-ready bispecific antibodies for high-throughput, biologically relevant assays. We identified an obligate bispecific co-targeting CD79a/b and CD22 as a potent inhibitor of human B cell activation from a short-term flow cytometry signaling assay. A long-term, high-content imaging assay identified anti-integrin bispecific inhibitors of human cell matrix accumulation targeting integrins β1 and β6 or αV and β1. In all cases, functional activity was conserved from the bispecific screening format to a therapeutically relevant format. We also introduce a broader type of mechanistic screen whereby functional modulation of different cell subsets in peripheral blood mononuclear cells was evaluated simultaneously. We identified bispecific antibodies capable of activating different T cell subsets of potential interest for applications in oncology or infectious disease, as well as bispecifics abrogating T cell activity of potential interest to autoimmune or inflammatory disease. The bispecific target pair discovery technology described herein offers access to new target biology and unique bispecific therapeutic opportunities in diverse disease indications. Taylor & Francis 2021-01-25 /pmc/articles/PMC7849716/ /pubmed/33487120 http://dx.doi.org/10.1080/19420862.2020.1859049 Text en © 2021 The Author(s). Published with license by Taylor & Francis Group, LLC. https://creativecommons.org/licenses/by-nc/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (http://creativecommons.org/licenses/by-nc/4.0/ (https://creativecommons.org/licenses/by-nc/4.0/) ), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Report
Bhatta, Pallavi
Whale, Kevin D.
Sawtell, Amy K.
Thompson, Clare L.
Rapecki, Stephen E.
Cook, David A.
Twomey, Breda M.
Mennecozzi, Milena
Starkie, Laura E.
Barry, Emily M. C.
Peters, Shirley J.
Kamal, Ahmad M.
Finney, Helene M.
Bispecific antibody target pair discovery by high-throughput phenotypic screening using in vitro combinatorial Fab libraries
title Bispecific antibody target pair discovery by high-throughput phenotypic screening using in vitro combinatorial Fab libraries
title_full Bispecific antibody target pair discovery by high-throughput phenotypic screening using in vitro combinatorial Fab libraries
title_fullStr Bispecific antibody target pair discovery by high-throughput phenotypic screening using in vitro combinatorial Fab libraries
title_full_unstemmed Bispecific antibody target pair discovery by high-throughput phenotypic screening using in vitro combinatorial Fab libraries
title_short Bispecific antibody target pair discovery by high-throughput phenotypic screening using in vitro combinatorial Fab libraries
title_sort bispecific antibody target pair discovery by high-throughput phenotypic screening using in vitro combinatorial fab libraries
topic Report
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7849716/
https://www.ncbi.nlm.nih.gov/pubmed/33487120
http://dx.doi.org/10.1080/19420862.2020.1859049
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