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目的采用表面等离子共振(surface plasmon resonance,SPR)技术快速检测痕量牛血清白蛋白(bovine serum albumin,BSA)含量。方法将兔抗BSA抗体标记到表面等离子共振芯片表面,监测BSA与抗体结合后SPR芯片表面的变化,利用SPR技术建立快速检测痕量BSA的新方法,对建立的方法进行检出限、定量限和最佳线性范围的确定,并进行重复性验证及初步应用。结果抗体的标记水平为10 000 RU,对BSA具有良好的亲和能力;基于信噪比为3时,BSA的检出限为0.003 ng/ml,基于信噪比为10时,BSA的定量限为0.01 ng/ml,BSA浓度在0.01~100 ng/ml范围内,与SPR仪响应值的变化量呈良好的线性关系,R2=0.997 9;用建立的SPR方法检测不同浓度BSA的日内RSD为3.3%~4.6%,日间RSD为3.4%~4.8%,均小于5%;SPR方法和间接竞争酶联免疫测定法(indirect competitive ELISA,icELISA)检测不同浓度BSA含量的结果比较,差异无统计学意义(P>0.05)。结论已建立了采用SPR技术检测BSA的新方法,可用于痕量BSA的快速检测。
Objective To rapidly detect the content of bovine serum albumin (BSA) by surface plasmon resonance (SPR). Methods Rabbit anti-BSA antibody was labeled onto the surface plasmon resonance chip surface. The surface of SPR chip after BSA binding with antibody was monitored. A new method for rapid detection of trace BSA was established by SPR technique. The detection limits and the limits of quantitation And the determination of the best linear range, and repeated verification and preliminary application. Results The antibody labeled level was 10 000 RU and had good affinity to BSA. The detection limit of BSA was 0.003 ng / ml when the signal to noise ratio was 3, and the limit of quantitation The concentration of BSA in the range of 0.01-100 ng / ml showed a good linear relationship with the change of SPR response value, R2 = 0.997 9. The RSD of different concentrations of BSA detected by SPR method was 3.3% -4.6%, daytime RSD was 3.4% -4.8%, both less than 5%; SPR method and indirect competitive ELISA (indirect competitive ELISA, icELISA) test results of different concentrations of BSA content, the difference was not statistically Significance (P> 0.05). Conclusion A new method for detecting BSA by SPR has been established, which can be used for the rapid detection of trace BSA.