Author/Authors :
Yousefi, Mohammadreza Department of Biotechnology - Higher Education Institute of Rab-Rashid, Tabriz, Iran , Safar Farajnia Drug Applied Research Center - Tabriz University of Medical Sciences, Tabriz, Iran , Mokhtarzadeh, Ahad Department of Biotechnology - Higher Education Institute of Rab-Rashid, Tabriz, Iran , Akbari, Bahman Department of Medical Biotechnology - Faculty of Medicine - Kermanshah University of Medical Sciences, Kermanshah, Iran , Ahdi Khosroshahi, Shiva Biotechnology Research Center - Tabriz University of Medical Sciences, Tabriz, Iran , Mamipour, Mina Department of Biotechnology - Higher Education Institute of Rab-Rashid, Tabriz, Iran , Dariushnejad, Hassan Immunology Research Center - Tabriz University of Medical Sciences, Tabriz, Iran , Ahmadzadeh, Vahideh Biotechnology Research Center - Tabriz University of Medical Sciences, Tabriz, Iran
Abstract :
Background: 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 Escherichia coli (E. coli) 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 E.
coli.
Methods: For this purpose, the fragment coding for anti-CD20 huscFv subcloned into
the pET22b (+) and transformed into the E. coli 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.
Result: 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 μg/ml soluble anti-CD20 scFv.
Conclusion: The result of this study demonstrated remarkable effect of pKJE7 chaperone
on enhancement of soluble expression of anti-CD20 huscFv antibody in E. coli.
