CAJ | 학술논문

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芯片比较在磺胺甲噁唑(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.
목적 연제일충신형적환원태우혈청백단백(rBSA)심편,사기괄용우Biacore계렬의기병구유여CM5심편상동적공능.방법 이용이류건환원제장천연태우혈청백단백(nBSA)적소유구기환원출래,형성rBSA,연후통과물리흡부장rBSA고정우라금심편표면,경교련화최기화처리후득도rBSA심편,접착이용원자력현미경(AFM)표정기표면수식효과,병여CM5심편비교재광알갑오서(SMX)검측중적응용효과.결과 통과AFM소묘,발현rBSA심편표면기질균일;기검측SMX적억제표준곡선여CM5심편상비교,곡선형상십분상사,각억제농도적항체결합신호균구유흔호적은정성;차외,rBSA심편환구유재생경용역、분석시간경단적우세.결론 rBSA심편수식성공,제비적심편구유우량적성능,능구만족실험적수요.
Objective To prepare a reduced bovine serum albumin (rBSA) sensor chip,which is a new sensor chip for the Biacore instrument and carl achieve the salve 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 earboxylation 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 inhibitive immunoassay format,which was the same method used in the SMX detection by the CM5 sensor chip in the previous study.Results The stnface of the rBSA sensor chip Was homogeneous as observed by AFM.The ealibration 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.