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
Descripción
Sumario: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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