高灵敏的蛋白质免疫印迹新方法用于β-淀粉样蛋白检测  

A High Sensitive Western Blotting Method for the Detection of β-Amyloid Protein

作　　者：冯爱平[1] 扎拉嘎胡[2] 李威[2] 刘英富[2] 

机构地区：[1]四川省德阳市第二人民医院神经外科,四川德阳61800 [2]武警后勤学院细胞生物学与医学遗传学教研室,天津300162

出　　处：《生物技术通讯》2014年第2期245-247,251,共4页Letters in Biotechnology

基　　金：武警后勤学院(原武警医学院)院级课题(WYB201116)

摘　　要：目的:对抗体进行改进,降低传统蛋白质免疫印迹方法的检出限。方法:在传统蛋白质免疫印迹方法的基础上,将一抗和二抗分别与纳米金和氧化石墨烯结合,以提高其对痕量样本的检测灵敏度,降低检测限。结果:优化后的方法对β-淀粉样蛋白的检测限从原来的432 pg/mL降低至16 pg/mL,为原来的1/27。结论:改进后的蛋白质免疫印迹新方法性能稳定,重复性好,可用于生物样本中痕量被检测物的测定。Objective： The antibodies were improved to reduce the limit of detection of traditional Western blot method. Methods： The method was used for sensitive detection of target protein based on the principle of specific binding between antigen and antibody. Gold nanoparticles and graphene oxide were introduced and combined with primary antibody and second antibody respectively in this study, and the aim was to lower detection limit. Re sults： The modified Western blotting method was able to detect β-amyloid protein with concentration of 16 pg/mL comparing with 432 pg/mL in traditional Western blotting method, and the detection limit was decreased to 1/27. Conclusion： The optimized method owns superior performance, good reproducibility, and can be used for the deter mination of trace analytes in biological samples.

关 键 词：蛋白质免疫印迹 Β-淀粉样蛋白 纳米金 氧化石墨烯 

分 类 号：Q502[生物学—生物化学]