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Enzyme-linked immunosorbent assays for insulin-like growth factor-I using six-histidine tag fused proteins

Yong Huang1, Ruina Shi, Xuefei Zhong

  • 1College of Chemistry and Molecular Engineering, Peking University, Beijing 100871, China.

Analytica Chimica Acta
|July 10, 2007
PubMed

The fusion proteins of insulin-like growth factor-I (IGF-I) and six-histidine tag (IGF-I-6H, 6H-IGF-I-6H) were cloned, expressed, purified and renatured, with their immunoreaction properties and biological activities intact. The binding kinetics between these fusion proteins and anti-IGF-I antibody or anti-6H antibody were studied using surface plasmon resonance (SPR). Two enzyme-linked immunosorbent assay (ELISA) modes, which proved feasible in the measurement of human serum samples, were used to detect IGF-I with the help of the six-histidine tagged proteins. Furthermore, combining the production technique of the six-histidine tagged fusion protein with the competitive sandwich ELISA mode, using an enzyme labeled anti-6H antibody as a tracer, can be a universal immunochemical method to quantitate other polypeptides or proteins.

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