摘要
目的 研制一种新型的还原态牛血清白蛋白（rBSA）芯片，使其适用于Biacore系列仪器并具有与CM5芯片相同的功能。方法 利用二硫键还原剂将天然态牛血清白蛋白（nBSA）的所有巯基还原出来，形成rBSA，然后通过物理吸附将rBSA固定于裸金芯片表面，经交联和羧基化处理后得到rBSA芯片，接着利用原子力显微镜（AFM）表征其表面修饰效果，并与CM5芯片比较在磺胺甲唾唑（SMX）检测中的应用效果。结果 通过AFM扫描，发现rBSA芯片表面基质均一；其检测SMX的抑制标准曲线与CM5芯片相比较，曲线形状十分相似，各抑制浓度的抗体结合信号均具有很好的稳定性；此外，rBSA芯片还具有再生更容易、分析时间更短的优势。结论 rBSA芯片修饰成功，制备的芯片具有优良的性能，能够满足实验的需要。
Objective To prepare a reduced bovine serum albumin （rBSA） sensor chip, which is a new sensor chip for the Biacore instrument and can achieve the same function as the CM5 sensor chip. Methods Native BSA was transformed to rBSA by treatment with a disulfide bond reducing agent. Then rBSA was immobilized on the gold surface of the sensor chip by physical adsorption. After subsequent cross-linking reaction and carboxylation process, the preparation of the rBSA sensor chip was achieved. The surface of the rBSA sensor chip was observed by atomic force microscopy（AFM）. The rBSA sensor chip was applied to detect sulfamethoxazole（SMX） using an inhib- itive immunoassay format, which was the same method used in the SMX detection by the CM5 sensor chip in the previous study. Results The surface of the rBSA sensor chip was homogeneous as observed by AFM. The calibration curve for the measurement of SMX using the rBSA sensor chip was similar to the one using the CM5 sensor chip, and the.binding of antibody to the rBSA sensor chip had good reproducibility. In addition, compared to the CM5 sensor chip, the rBSA sensor chip had the advantage of easier regeneration and shorter analytical time in the detection of SMX. Conclusion The new rBSA sensor chip was successfully prepared. It was demonstrated that the rBSA sensor chip had excellent performance in the determination of SMX.
出处
《国际生物医学工程杂志》
CAS
北大核心
2009年第1期5-8,共4页
International Journal of Biomedical Engineering
基金
国家“863”计划资助项目（2006AA062406）
国家自然科学基金资助项目（50830106,30727001）
