Expression and Purification of scFv(2H7)-P18F3, a Bi-Modular Fusion Protein (BMFP) Targeting Human CD20

P18F3-based bi-modular fusion proteins (BMFPs), designed to re-direct pre-existing anti-Epstein-Barr virus (EBV) endogenous polyclonal antibodies towards defined target cells, demonstrated efficient biological activity in a mouse tumor model and could potentially represent a universal and versatile...

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Detalles Bibliográficos
Autores principales: Brousse, Carine, Rainey, Nathan E., Desrames, Alexandra, Teillaud, Jean-Luc, Gamain, Benoît, Chêne, Arnaud
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Bio-Protocol 2023
Materias:
Methods Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10213076/
https://www.ncbi.nlm.nih.gov/pubmed/37251097
http://dx.doi.org/10.21769/BioProtoc.4682
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author Brousse, Carine
Rainey, Nathan E.
Desrames, Alexandra
Teillaud, Jean-Luc
Gamain, Benoît
Chêne, Arnaud
author_facet Brousse, Carine
Rainey, Nathan E.
Desrames, Alexandra
Teillaud, Jean-Luc
Gamain, Benoît
Chêne, Arnaud
author_sort Brousse, Carine
collection PubMed
description P18F3-based bi-modular fusion proteins (BMFPs), designed to re-direct pre-existing anti-Epstein-Barr virus (EBV) endogenous polyclonal antibodies towards defined target cells, demonstrated efficient biological activity in a mouse tumor model and could potentially represent a universal and versatile platform to develop novel therapeutics against a broad range of diseases. This protocol provides step-by-step instructions for expressing scFv(2H7)-P18F3, a BMFP targeting human CD20, in Escherichia coli (SHuffle(®)), and for purifying soluble proteins using a two-step process, namely immobilized metal affinity chromatography (IMAC) followed by size exclusion chromatography. This protocol can also be used for expression and purification of other BMFPs with alternative binding specificities.
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spelling pubmed-102130762023-05-27 Expression and Purification of scFv(2H7)-P18F3, a Bi-Modular Fusion Protein (BMFP) Targeting Human CD20 Brousse, Carine Rainey, Nathan E. Desrames, Alexandra Teillaud, Jean-Luc Gamain, Benoît Chêne, Arnaud Bio Protoc Methods Article P18F3-based bi-modular fusion proteins (BMFPs), designed to re-direct pre-existing anti-Epstein-Barr virus (EBV) endogenous polyclonal antibodies towards defined target cells, demonstrated efficient biological activity in a mouse tumor model and could potentially represent a universal and versatile platform to develop novel therapeutics against a broad range of diseases. This protocol provides step-by-step instructions for expressing scFv(2H7)-P18F3, a BMFP targeting human CD20, in Escherichia coli (SHuffle(®)), and for purifying soluble proteins using a two-step process, namely immobilized metal affinity chromatography (IMAC) followed by size exclusion chromatography. This protocol can also be used for expression and purification of other BMFPs with alternative binding specificities. Bio-Protocol 2023-05-20 /pmc/articles/PMC10213076/ /pubmed/37251097 http://dx.doi.org/10.21769/BioProtoc.4682 Text en Copyright © 2023 The Authors; exclusive licensee Bio-protocol LLC. https://creativecommons.org/licenses/by-nc/4.0/This is an open access article under the CC BY-NC license (https://creativecommons.org/licenses/by-nc/4.0/).
spellingShingle Methods Article
Brousse, Carine
Rainey, Nathan E.
Desrames, Alexandra
Teillaud, Jean-Luc
Gamain, Benoît
Chêne, Arnaud
Expression and Purification of scFv(2H7)-P18F3, a Bi-Modular Fusion Protein (BMFP) Targeting Human CD20
title Expression and Purification of scFv(2H7)-P18F3, a Bi-Modular Fusion Protein (BMFP) Targeting Human CD20
title_full Expression and Purification of scFv(2H7)-P18F3, a Bi-Modular Fusion Protein (BMFP) Targeting Human CD20
title_fullStr Expression and Purification of scFv(2H7)-P18F3, a Bi-Modular Fusion Protein (BMFP) Targeting Human CD20
title_full_unstemmed Expression and Purification of scFv(2H7)-P18F3, a Bi-Modular Fusion Protein (BMFP) Targeting Human CD20
title_short Expression and Purification of scFv(2H7)-P18F3, a Bi-Modular Fusion Protein (BMFP) Targeting Human CD20
title_sort expression and purification of scfv(2h7)-p18f3, a bi-modular fusion protein (bmfp) targeting human cd20
topic Methods Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10213076/
https://www.ncbi.nlm.nih.gov/pubmed/37251097
http://dx.doi.org/10.21769/BioProtoc.4682
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