AFP-L3蛋白芯片的建立与应用

目的:建立一种可同时检测血清中甲胎蛋白(AFP)及甲胎蛋白异质体(AFP-L3)的蛋白芯片方法,为AFP-L3检测提供经济、便捷、省时、有效的新途径。方法:将鼠源AFP单克隆抗体和小扁豆凝集素点样固定在醛基玻片上,制备出AFP抗体和小扁豆凝集素蛋白芯片。利用抗原抗体特异性结合以及盐藻糖与小扁豆凝集素特异性结合的原理,用蛋白芯片方法检测血清样本中的AFP和AFP-L3。结果:肝癌血清39份,其中37份检测到AFP;26份肝癌血清中同时检测到AFP和AFP-L3,2份肝癌血清均未检测到AFP和AFP-L3。肝细胞癌组血清样本中AFP和AFP-L3水平明显高于健康对照组(P0.001),健康对照组与空白对照组统计学上无差异(P0.05)。结论:本研究成功地建立AFP和AFP-L3同时检测的蛋白芯片方法,与ELISA法和凝集素微量离心柱法相比,是一种切实可行,经济、便捷、省时、有效的方法。

Development and Application of Protein Microarray for Detecting Alpha-fetoprotein-L3

ABSTRACT Objective: To develop a protein microarray method for detecting AFP and AFP-L3 in serum samples at the same time and provide an economical, feasible, time-saving and efficiency method for screening of primary hepatic carcinoma. Methods: Spotting mouse-derived AFP monoclonal antibody and Lens culinaris agglutinin on an aldehyde glass slide to develop a protein microarray method for detecting AFP and AFP-L3 of HCC patients and healthy controls. The protein microarray was based on the antibody-antigen reaction principle and Lens culinaris agglutinin-Fucose reaction principle. Results: AFP was detected in 37 cases of 39 HCC samples. AFP and AFP-L3 were both detected in 26 cases of 37 HCC samples. AFP and AFP-L3 were both undetected in 2 cases of 39 HCC sam- ples. The AFP and AFP-L3 levels in serum of HCC patients were significantly higher than those of healthy controls(P<0.001). There was no statistically significant difference between healthy controls and blank controls (P>0.05). Conclusion: A protein microarray which can assay AFP and AFP-L3 at same time was successfully established. Compared with Lens culinaris agglutinin-coupled spin column(ACSC) technology and ELISA, it was a feasible, economical, convenient use, time-saving and efficiency method.