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KMID : 0380020090240060591
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Korean Journal of Biotechnology and Bioengineering
2009 Volume.24 No. 6 p.591 ~ p.597
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Functional Expression of Anti-BNP scFv in E. coli Cytoplasm for the Detection of B-type Natriuretic Peptide
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Maeng Bo-Hee
Nam Dong-Hyun
Kim Yong-Hwan
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Abstract
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B-type natriuretic peptide is a neurohormone secreted in the cardiac ventricles. BNP levels are elevated in patients with ventricular dysfunction. Therefore, the concentration of BNP is important factor to reflect diagnosis and prognosis for cardiovascular disease. In this respect, anti-BNP scFv is an urgent requirement for early diagnosis in the field of biosensor. Herein, 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. Additionally, being exposed to both BNP and ANP, anti-BNP scFv specifically captured only BNP. Therefore, anti-BNP scFv expressed in this study will be applied to measure the concentration of BNP as a diagnostic recognition molecule.
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KEYWORD
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BNP, anti-BNP, scFv
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