earticle

영어

B-type natriuretic peptide is a cardiac neurohormone secreted from the ventricles in volume expansion and pressure overload. B-type natriuretic peptide levels are elevated in patients with left ventricular dysfunction and correlated to the severity as well as prognosis. Therefore, B-type
natriuretic peptide is a potential biomarker as well as a therapeutic target. Herein, we report the expression of anti-BNP scFv in the cytoplasm of Escherichia coli for the detection of B-type natriuretic peptide. In this study, we investigated optimal vector, temperature and IPTG concentration for expression of anti-BNP scFv and compare affinity for BNP with ANP. The genetic codes of anti-BNP scFv were chemically synthesized and cloned into both pET22b(+) and pColdⅣ vector, respectively. The recombinant scFv was successfully expressed as a functional form in cytoplasm of E. coli and detected through Western blot and ELISA. The
highest level of functional expression of anti-BNP scFv was achieved using pET22b(+) vector at 15℃ by addition of 0.1 mM IPTG. And we confirm expressed anti-BNP scFv specifically captured only BNP. Additionally, we also examined the effect of molecular chaperones on the expression level of functional anti-BNP scFv by coexpressing chaperone gene with the scFv. It was found out that co-expression of groES-groEL chaperon plasmid increased the scFv soluble expression
level and activity compared to the scFv expressed without chaperone coexpression.