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Novel Selection Approaches to Identify Antibodies Targeting Neoepitopes on the C5b6 Intermediate Complex to Inhibit Membrane Attack Complex Formation

The terminal pathway of complement is implicated in the pathology of multiple diseases and its inhibition is, therefore, an attractive therapeutic proposition. The practicalities of inhibiting this pathway, however, are challenging, as highlighted by the very few molecules in the clinic. The protein...

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Autores principales: Stach, Lasse, Dinley, Emily K. H., Tournier, Nadia, Bingham, Ryan P., Gormley, Darren A., Bramhall, Jo L., Taylor, Adam, Clarkson, Jane E., Welbeck, Katherine A., Harris, Claire L., Feeney, Maria, Hughes, Jane P., Sepp, Armin, Batuwangala, Thil D., Kitchen, Semra J., Nichols, Eva-Maria
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8544208/
https://www.ncbi.nlm.nih.gov/pubmed/34698051
http://dx.doi.org/10.3390/antib10040039
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author Stach, Lasse
Dinley, Emily K. H.
Tournier, Nadia
Bingham, Ryan P.
Gormley, Darren A.
Bramhall, Jo L.
Taylor, Adam
Clarkson, Jane E.
Welbeck, Katherine A.
Harris, Claire L.
Feeney, Maria
Hughes, Jane P.
Sepp, Armin
Batuwangala, Thil D.
Kitchen, Semra J.
Nichols, Eva-Maria
author_facet Stach, Lasse
Dinley, Emily K. H.
Tournier, Nadia
Bingham, Ryan P.
Gormley, Darren A.
Bramhall, Jo L.
Taylor, Adam
Clarkson, Jane E.
Welbeck, Katherine A.
Harris, Claire L.
Feeney, Maria
Hughes, Jane P.
Sepp, Armin
Batuwangala, Thil D.
Kitchen, Semra J.
Nichols, Eva-Maria
author_sort Stach, Lasse
collection PubMed
description The terminal pathway of complement is implicated in the pathology of multiple diseases and its inhibition is, therefore, an attractive therapeutic proposition. The practicalities of inhibiting this pathway, however, are challenging, as highlighted by the very few molecules in the clinic. The proteins are highly abundant, and assembly is mediated by high-affinity protein–protein interactions. One strategy is to target neoepitopes that are present transiently and only exist on active or intermediate complexes but not on the abundant native proteins. Here, we describe an antibody discovery campaign that generated neoepitope-specific mAbs against the C5b6 complex, a stable intermediate complex in terminal complement complex assembly. We used a highly diverse yeast-based antibody library of fully human IgGs to screen against soluble C5b6 antigen and successfully identified C5b6 neoepitope-specific antibodies. These antibodies were diverse, showed good binding to C5b6, and inhibited membrane attack complex (MAC) formation in a solution-based assay. However, when tested in a more physiologically relevant membrane-based assay these antibodies failed to inhibit MAC formation. Our data highlight the feasibility of identifying neoepitope binding mAbs, but also the technical challenges associated with the identification of functionally relevant, neoepitope-specific inhibitors of the terminal pathway.
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spelling pubmed-85442082021-10-26 Novel Selection Approaches to Identify Antibodies Targeting Neoepitopes on the C5b6 Intermediate Complex to Inhibit Membrane Attack Complex Formation Stach, Lasse Dinley, Emily K. H. Tournier, Nadia Bingham, Ryan P. Gormley, Darren A. Bramhall, Jo L. Taylor, Adam Clarkson, Jane E. Welbeck, Katherine A. Harris, Claire L. Feeney, Maria Hughes, Jane P. Sepp, Armin Batuwangala, Thil D. Kitchen, Semra J. Nichols, Eva-Maria Antibodies (Basel) Article The terminal pathway of complement is implicated in the pathology of multiple diseases and its inhibition is, therefore, an attractive therapeutic proposition. The practicalities of inhibiting this pathway, however, are challenging, as highlighted by the very few molecules in the clinic. The proteins are highly abundant, and assembly is mediated by high-affinity protein–protein interactions. One strategy is to target neoepitopes that are present transiently and only exist on active or intermediate complexes but not on the abundant native proteins. Here, we describe an antibody discovery campaign that generated neoepitope-specific mAbs against the C5b6 complex, a stable intermediate complex in terminal complement complex assembly. We used a highly diverse yeast-based antibody library of fully human IgGs to screen against soluble C5b6 antigen and successfully identified C5b6 neoepitope-specific antibodies. These antibodies were diverse, showed good binding to C5b6, and inhibited membrane attack complex (MAC) formation in a solution-based assay. However, when tested in a more physiologically relevant membrane-based assay these antibodies failed to inhibit MAC formation. Our data highlight the feasibility of identifying neoepitope binding mAbs, but also the technical challenges associated with the identification of functionally relevant, neoepitope-specific inhibitors of the terminal pathway. MDPI 2021-10-12 /pmc/articles/PMC8544208/ /pubmed/34698051 http://dx.doi.org/10.3390/antib10040039 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Stach, Lasse
Dinley, Emily K. H.
Tournier, Nadia
Bingham, Ryan P.
Gormley, Darren A.
Bramhall, Jo L.
Taylor, Adam
Clarkson, Jane E.
Welbeck, Katherine A.
Harris, Claire L.
Feeney, Maria
Hughes, Jane P.
Sepp, Armin
Batuwangala, Thil D.
Kitchen, Semra J.
Nichols, Eva-Maria
Novel Selection Approaches to Identify Antibodies Targeting Neoepitopes on the C5b6 Intermediate Complex to Inhibit Membrane Attack Complex Formation
title Novel Selection Approaches to Identify Antibodies Targeting Neoepitopes on the C5b6 Intermediate Complex to Inhibit Membrane Attack Complex Formation
title_full Novel Selection Approaches to Identify Antibodies Targeting Neoepitopes on the C5b6 Intermediate Complex to Inhibit Membrane Attack Complex Formation
title_fullStr Novel Selection Approaches to Identify Antibodies Targeting Neoepitopes on the C5b6 Intermediate Complex to Inhibit Membrane Attack Complex Formation
title_full_unstemmed Novel Selection Approaches to Identify Antibodies Targeting Neoepitopes on the C5b6 Intermediate Complex to Inhibit Membrane Attack Complex Formation
title_short Novel Selection Approaches to Identify Antibodies Targeting Neoepitopes on the C5b6 Intermediate Complex to Inhibit Membrane Attack Complex Formation
title_sort novel selection approaches to identify antibodies targeting neoepitopes on the c5b6 intermediate complex to inhibit membrane attack complex formation
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8544208/
https://www.ncbi.nlm.nih.gov/pubmed/34698051
http://dx.doi.org/10.3390/antib10040039
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