Sensitive detection of HSP70 using a current-amplified biosensor based on antibody-loaded PS-AuNPs@Cys/Au modified ITO chip.

Abstract

A biosensing electrochemical platform for heat shock protein 70 (HSP70) has been developed by integrating a three-electrode indium tin oxide (ITO) on a chip. The platform includes modifications to the reference electrode and working electrode for the detection of HSP70. The new platform is constructed by assembly of HSP70 antibody on PS-AuNPs@Cys/Au indium tin oxide (ITO) electrode to create a high HSP70 sensitive surface. The PS-AuNPs@Cys/Au indium tin oxide (ITO) electrode is obtained by immersing the ITO electrode into the PS-AuNPs@Cys solution and performing constant potential deposition at -1.4V (Ag/AgCl). The PS-AuNPs@Cys/Au film deposited on ITO glass provides a desirable substrate for the immobilization of the HSP70 antibody and improves the loading of antibody between PS-AuNPs@Cys/Au and the electrode resulting in a significant amplification. Under optimal conditions, the fabricated sensor demonstrates a linear range extending from 0.1 ng mL- 1 to 1000 ng mL- 1, with an impressive detection limit of 25.7 pg mL- 1 (S/N = 3). The developed immunoassay method successfully detected the HSP70 content in normal human blood samples and outperformed the ELISA method commonly used for clinical sample analysis.