Expression and Purification of scFv2H7-P18F3, a Bi-Modular Fusion Protein (BMFP) Targeting Human CD20

Carine Brousse; Nathan E Rainey; Alexandra Desrames; Jean-Luc Teillaud; Benoît Gamain; Arnaud Chêne

doi:10.21769/BioProtoc.4682

Expression and Purification of scFv_2H7-P18F3, a Bi-Modular Fusion Protein (BMFP) Targeting Human CD20

Bio Protoc. 2023 May 20;13(10):e4682. doi: 10.21769/BioProtoc.4682.

Authors

Carine Brousse¹, Nathan E Rainey¹, Alexandra Desrames¹, Jean-Luc Teillaud², Benoît Gamain¹, Arnaud Chêne¹

Affiliations

¹ Université Paris Cité and Université des Antilles, INSERM, BIGR, F-75015 Paris, France.
² Laboratoire «Microenvironnement Immunitaire et Immunothérapie», Inserm U.1135, Centre d'Immunologie et des Maladies Infectieuses (CIMI-Paris), Faculté de médecine, Sorbonne Université, Paris, France.

Abstract

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.

Keywords: Anti-EBV antibodies; B lymphoma; BMFP; CD20; Escherichia coli SHuffle®; Fusion protein; Immunotherapy; scFv.