Cargando…

Evaluation of Phage Display Biopanning Strategies for the Selection of Anti-Cell Surface Receptor Antibodies

Monoclonal antibodies (mAbs) are one of the most successful and versatile protein-based pharmaceutical products used to treat multiple pathological conditions. The remarkable specificity of mAbs and their affinity for biological targets has led to the implementation of mAbs in the therapeutic regime...

Descripción completa

Detalles Bibliográficos
Autores principales: Panagides, Nadya, Zacchi, Lucia F., De Souza, Mitchell J., Morales, Rodrigo A. V., Karnowski, Alexander, Liddament, Mark T., Owczarek, Catherine M., Mahler, Stephen M., Panousis, Con, Jones, Martina L., Fercher, Christian
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9369378/
https://www.ncbi.nlm.nih.gov/pubmed/35955604
http://dx.doi.org/10.3390/ijms23158470
_version_ 1784766437898846208
author Panagides, Nadya
Zacchi, Lucia F.
De Souza, Mitchell J.
Morales, Rodrigo A. V.
Karnowski, Alexander
Liddament, Mark T.
Owczarek, Catherine M.
Mahler, Stephen M.
Panousis, Con
Jones, Martina L.
Fercher, Christian
author_facet Panagides, Nadya
Zacchi, Lucia F.
De Souza, Mitchell J.
Morales, Rodrigo A. V.
Karnowski, Alexander
Liddament, Mark T.
Owczarek, Catherine M.
Mahler, Stephen M.
Panousis, Con
Jones, Martina L.
Fercher, Christian
author_sort Panagides, Nadya
collection PubMed
description Monoclonal antibodies (mAbs) are one of the most successful and versatile protein-based pharmaceutical products used to treat multiple pathological conditions. The remarkable specificity of mAbs and their affinity for biological targets has led to the implementation of mAbs in the therapeutic regime of oncogenic, chronic inflammatory, cardiovascular, and infectious diseases. Thus, the discovery of novel mAbs with defined functional activities is of crucial importance to expand our ability to address current and future clinical challenges. In vitro, antigen-driven affinity selection employing phage display biopanning is a commonly used technique to isolate mAbs. The success of biopanning is dependent on the quality and the presentation format of the antigen, which is critical when isolating mAbs against membrane protein targets. Here, we provide a comprehensive investigation of two established panning strategies, surface-tethering of a recombinant extracellular domain and cell-based biopanning, to examine the impact of antigen presentation on selection outcomes with regards to the isolation of positive mAbs with functional potential against a proof-of-concept type I cell surface receptor. Based on the higher sequence diversity of the resulting antibody repertoire, presentation of a type I membrane protein in soluble form was more advantageous over presentation in cell-based format. Our results will contribute to inform and guide future antibody discovery campaigns against cell surface proteins.
format Online
Article
Text
id pubmed-9369378
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-93693782022-08-12 Evaluation of Phage Display Biopanning Strategies for the Selection of Anti-Cell Surface Receptor Antibodies Panagides, Nadya Zacchi, Lucia F. De Souza, Mitchell J. Morales, Rodrigo A. V. Karnowski, Alexander Liddament, Mark T. Owczarek, Catherine M. Mahler, Stephen M. Panousis, Con Jones, Martina L. Fercher, Christian Int J Mol Sci Article Monoclonal antibodies (mAbs) are one of the most successful and versatile protein-based pharmaceutical products used to treat multiple pathological conditions. The remarkable specificity of mAbs and their affinity for biological targets has led to the implementation of mAbs in the therapeutic regime of oncogenic, chronic inflammatory, cardiovascular, and infectious diseases. Thus, the discovery of novel mAbs with defined functional activities is of crucial importance to expand our ability to address current and future clinical challenges. In vitro, antigen-driven affinity selection employing phage display biopanning is a commonly used technique to isolate mAbs. The success of biopanning is dependent on the quality and the presentation format of the antigen, which is critical when isolating mAbs against membrane protein targets. Here, we provide a comprehensive investigation of two established panning strategies, surface-tethering of a recombinant extracellular domain and cell-based biopanning, to examine the impact of antigen presentation on selection outcomes with regards to the isolation of positive mAbs with functional potential against a proof-of-concept type I cell surface receptor. Based on the higher sequence diversity of the resulting antibody repertoire, presentation of a type I membrane protein in soluble form was more advantageous over presentation in cell-based format. Our results will contribute to inform and guide future antibody discovery campaigns against cell surface proteins. MDPI 2022-07-30 /pmc/articles/PMC9369378/ /pubmed/35955604 http://dx.doi.org/10.3390/ijms23158470 Text en © 2022 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
Panagides, Nadya
Zacchi, Lucia F.
De Souza, Mitchell J.
Morales, Rodrigo A. V.
Karnowski, Alexander
Liddament, Mark T.
Owczarek, Catherine M.
Mahler, Stephen M.
Panousis, Con
Jones, Martina L.
Fercher, Christian
Evaluation of Phage Display Biopanning Strategies for the Selection of Anti-Cell Surface Receptor Antibodies
title Evaluation of Phage Display Biopanning Strategies for the Selection of Anti-Cell Surface Receptor Antibodies
title_full Evaluation of Phage Display Biopanning Strategies for the Selection of Anti-Cell Surface Receptor Antibodies
title_fullStr Evaluation of Phage Display Biopanning Strategies for the Selection of Anti-Cell Surface Receptor Antibodies
title_full_unstemmed Evaluation of Phage Display Biopanning Strategies for the Selection of Anti-Cell Surface Receptor Antibodies
title_short Evaluation of Phage Display Biopanning Strategies for the Selection of Anti-Cell Surface Receptor Antibodies
title_sort evaluation of phage display biopanning strategies for the selection of anti-cell surface receptor antibodies
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9369378/
https://www.ncbi.nlm.nih.gov/pubmed/35955604
http://dx.doi.org/10.3390/ijms23158470
work_keys_str_mv AT panagidesnadya evaluationofphagedisplaybiopanningstrategiesfortheselectionofanticellsurfacereceptorantibodies
AT zacchiluciaf evaluationofphagedisplaybiopanningstrategiesfortheselectionofanticellsurfacereceptorantibodies
AT desouzamitchellj evaluationofphagedisplaybiopanningstrategiesfortheselectionofanticellsurfacereceptorantibodies
AT moralesrodrigoav evaluationofphagedisplaybiopanningstrategiesfortheselectionofanticellsurfacereceptorantibodies
AT karnowskialexander evaluationofphagedisplaybiopanningstrategiesfortheselectionofanticellsurfacereceptorantibodies
AT liddamentmarkt evaluationofphagedisplaybiopanningstrategiesfortheselectionofanticellsurfacereceptorantibodies
AT owczarekcatherinem evaluationofphagedisplaybiopanningstrategiesfortheselectionofanticellsurfacereceptorantibodies
AT mahlerstephenm evaluationofphagedisplaybiopanningstrategiesfortheselectionofanticellsurfacereceptorantibodies
AT panousiscon evaluationofphagedisplaybiopanningstrategiesfortheselectionofanticellsurfacereceptorantibodies
AT jonesmartinal evaluationofphagedisplaybiopanningstrategiesfortheselectionofanticellsurfacereceptorantibodies
AT fercherchristian evaluationofphagedisplaybiopanningstrategiesfortheselectionofanticellsurfacereceptorantibodies