摘要
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 determine trace bovine serum albumin（BSA）by surface plasmon resonance technology（SPR）. Methods SPR method was developed by monitoring the interaction between BSA and labeled rabbit anti-BSA antibody on the SPR sensor chip,then determined for detection limit,quantitative detection limit and optimal linear range, verified for reproducibility and applied preliminarily. Results The rabbit anti-BSA antibody at a labeling level of 10 000 RU showed good affinity to BSA. The detection limit of BSA based on a signal-to-noise（S / N）ratio of 3 was 0. 003 ng / ml, while that based on a S / N ratio of 10 was 0. 01 ng / ml. The BSA concentration at a range of 0. 01- 100 ng / ml showed good linear relationship to the response value of SPR analyzer,with a R2value of 0. 997 9. The RSDs of intra- and inter- assays on BSA at various concentrations by the developed SPR method were 3. 3% - 4. 6% and 3. 4% - 4. 8% respectively,both of which were less than 5%. No significant differences were observed between the determination results of BSA at various concentrations by SPR method and by indirect competitive ELISA（icELISA）（P〉 0. 05）. Conclusion A new SPR method was developed,which might be used for rapid determination of trace BSA.
出处
《中国生物制品学杂志》
CAS
CSCD
2014年第2期254-256,261,共4页
Chinese Journal of Biologicals
基金
国家自然科学基金资助项目(21277054)
关键词
表面等离子共振技术
牛血清白蛋白
含量测定
抗体
Surface plasmon resonance（SPR） technology
Bovine serum albumin（BSA）
Content determination
Antibody
