5,5'-Dithiobis(succinimidyl-2-nitrobenzoate) Functionalized Surface-enhanced Raman Scattering Nanoprobe for Immunoassay†

doi:10.7503/cjcu20150116

Abstract

Abstract:

A biofunctionalized nanoprobe for surface-enhanced Raman scattering(SERS)-based immunoassay was designed. 5,5'-Dithiobis(succinimidyl-2-nitrobenzoate)(DSNB) was employed as a linker for protein and gold nanoparticle, which can help to maintain the biological activity of the proteins. Besides that, DSNB possesses a high Raman scattering section, which can be used as the SERS probe as quantitative determination of proteins. Two kinds of biological systems(avidin-streptavidin and antigen-antibody) were selected for the target protein and nanoprobe specific binding to the protein chip. Meanwhile, silicon slide was employed as a carrier, which provides a Raman peak at 520 cm^-1 and can be used as a good internal standard for quantitative analysis of proteins. Using this method, anti-human IgG concentration in solutions can be quantitatively measured at concentrations as low as 5 pg/mL.

Key words: Surface-enhanced Raman scattering(SERS), Immunoassay, Nanoprobe, Avidin, Streptavidin

CLC Number:

O657

TrendMD:

CHEN Lei, XUE Xiangxin, CUI Yuncheng, HAN Xiaoxia, XU Weiqing, ZHAO Bing. 5,5'-Dithiobis(succinimidyl-2-nitrobenzoate) Functionalized Surface-enhanced Raman Scattering Nanoprobe for Immunoassay^†[J]. Chem. J. Chinese Universities, 2015, 36(8): 1505.

Figures/Tables 7

Scheme 1 Preparation of gold nanoprobe

Fig.1 Proposed biofunctional gold NPs for immunoassay

Fig.2 TEM image of gold NPs covered with DSNB and streptavidin(A) and SEM image of aggregated gold NPs after immunoassay(B)

Fig.3 UV-Vis spectra of DSNB(1×10-4 mol/L)(a), DSNB-streptavidin(100 μg/mL)(b), gold nano-particles covered with DSNB and streptavidin(c), gold nanoparticles(d) and gold nanoparticles covered with DSNB(e)

Fig.4 SERS spectra of DSNB before(a) and after(b—j) immunoassay with different concentrations of anti-human IgGConcentrations of anti-human IgG from b to j: 1×10-5, 5×10-6, 1×10-6, 5×10-7, 1×10-7, 5×10-8, 1×10-8, 5×10-9 and 0 mg/mL, respectively.

Fig.5 Concentration-dependent SERS intensities at 1336 cm-1 for DSNB containing different concentrations of anti-human IgG

Fig.6 Protein-induced SERS intensity changes of the DSNB-functionalized nanoprobe for immunoassay of anti-human IgG(5×10-7 mg/mL) detection systemFBS including 3×10-7—4×10-7 mg/mL protein; BSA: 5×10-7mg/mL; HSA: 5×10-7 mg/mL; Lys: 5×10-7 mg/mL.

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