Soluble Expression of Humanized Anti-CD20 Single Chain Antibody in Escherichia coli by Cytoplasmic Chaperones Co-expression

CD20 is an important cell surface receptor that is used for target therapy of B cell lymphoma and some related blood diseases due to vital function of CD20. In previous studies, a Rituximab based humanized single chain variable fragment (scFv) antibody showed good reactivity against B cell related c...

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Bibliographic Details
Published in:Avicenna journal of medical biotechnology 2018-07, Vol.10 (3), p.141-146
Main Authors: Yousefi, Mohammadreza, Farajnia, Safar, Mokhtarzadeh, Ahad, Akbari, Bahman, Ahdi Khosroshahi, Shiva, Mamipour, Mina, Dariushnejad, Hassan, Ahmadzadeh, Vahideh
Format: Article
Language:English
Subjects:
Antibodies
Cancer cells
Care and treatment
Escherichia coli
Lymphomas
Original
Viral antibodies
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Summary:CD20 is an important cell surface receptor that is used for target therapy of B cell lymphoma and some related blood diseases due to vital function of CD20. In previous studies, a Rituximab based humanized single chain variable fragment (scFv) antibody showed good reactivity against B cell related cancer cells. But this recombinant protein produced Inclusion Bodies (IBs) in ) cytoplasm. The aim of this study was to investigate the effect of coexpression with cytoplasmic chaperones on expression and solubility of humanized anti-CD20 scFv in . For this purpose, the fragment coding for anti-CD20 huscFv subcloned into the pET22b (+) and transformed into the BL21 (DE3) was evaluated. In order to inhibit the production of IBs, the effects of co-expression with cytoplasmic chaperones GroEL, DnaK, GroES, Tig, DnaJ and GrpE were investigated. Coexpression with cytoplasmic chaperones led to increased soluble expression of anti-CD20 recombinant protein. Among investigated chaperones, pKJE7 chaperone plasmid containing DnaJ, GrpE, DnaK chaperone genes had significant effects with an expression yield of 325 soluble anti-CD20 scFv. The result of this study demonstrated remarkable effect of pKJE7 chaperone on enhancement of soluble expression of anti-CD20 huscFv antibody in .
ISSN:2008-2835
2008-4625