TY  - JOUR
TI  - Expression and Purification of scFv2H7-P18F3, a Bi-Modular Fusion Protein (BMFP) Targeting Human CD20
AU  - Brousse, Carine 
AU  - Rainey, Nathan E.
AU  - Desrames, Alexandra 
AU  - Teillaud, Jean- Luc
AU  - Gamain, Benoît 
AU  - Chêne, Arnaud 
VL  - 13
IS  - 10
PY  - 2023
DA  - 2023/05/20
SP  - e4682
C1  - Bio-protocol 2023;13:e4682
DO  - 10.21769/BioProtoc.4682
UR  - https://doi.org/10.21769/BioProtoc.4682
AB  - 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 scFv2H7-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.
KW  - BMFP
KW  - Anti-EBV antibodies
KW  - Fusion protein
KW  - Immunotherapy
KW  - CD20
KW  - scFv
KW  - B lymphoma
KW  - Escherichia coli SHuffle
JF  - Bio-protocol
SN  - 2331-8325
PB  - Bio-protocol LLC.
BIO101  - False