琼脂糖自组装膜基底型免疫芯片载体制备及抗体固定研究  

Preparation of immune microarray carrier based on agarose self-assembled membrane and its application in probe immobilization

作　　者：马莉萍[1] 马生龙[1] 李云霞[1] 聂莹莹[1] 韩根亮[1] MA Li-Ping;MA Sheng-Long;LI Yun-Xia;NIE Ying- Ying;HAN Gen-Liang(Gansu Province Key Laboratory of Sensors and Sensing Technology, Institute of Sensor Technology, Gansu Academy of Sciences, Lanzhou 730000, Chin)

机构地区：[1]甘肃省科学院传感技术研究所甘肃省传感器与传感技术重点实验室,兰州730000

出　　处：《中国免疫学杂志》2018年第6期872-876,共5页Chinese Journal of Immunology

基　　金：甘肃省自然基金项目(1506RJZA191);甘肃省科学院应用开发项目(2016JK-05);甘肃省科学院院地合作项目(2014-CG-25);甘肃省科学院应用开发项目(2016JK-07);甘肃省科学院院地合作项目(2016-HZ-03)

摘　　要：目的:建立具有三维水凝胶结构的琼脂糖自组装膜载体的制备方法,提高免疫芯片对抗体的固定效率。方法:以玻片为载体,采用琼脂糖、高碘酸钠对玻片表面进行修饰,制备琼脂糖自组装膜载体。利用SEM、AFM和FTIR对载体进行表征,获得最佳制备条件。利用荧光显微成像及Image J软件,研究该载体对2种不同种属来源抗体的固定效率,比较琼脂糖自组装膜载体和普通醛基修饰载体对抗体的固定效果。结果:制备的琼脂糖自组装膜载体表面均匀、致密,这种结构比表面积大、抗体固定效率高。制备载体的最佳琼脂糖浓度为1.0%,其固定抗体的最佳浓度为0.3~0.4 mg/ml,琼脂糖自组装膜载体的荧光强度是普通醛基载体的3.94倍。结论:琼脂糖自组装膜是一种理想的固定抗体分子的表面修饰方法,有望用于免疫芯片载体的制备。Objective：To improve the immobilization efficiency of antibody molecules on immune microarray,the method of establishment and optimization of agarose self-assembled membrane carrier with three-dimensional hydrogel structure was established.Methods：The agarose self-assembled membrane carrier was prepared by using glass slide as the carrier,using agarose and sodium periodate modification on glass surface.The agarose self-assembled membrane carrier was characterized by TEM,AFM and FTIR.The optimum preparation conditions were obtained.The carrier for two different species of fixed source antibody efficiency were studied.Antibody loading capacity of agarose self-assembled membrane carrier and ordinary aldehyde carrier were investigated and compared by fluorescence microscopy imaging and Image J software.Results：The agarose nano-membrane carrier had uniform and compact surface.This structure could increase the specific surface area and improve the probe fixed rate.The optimal concentration of agarose for preparation of carrier was 1.0%.When the concentration of Ig G was 0.3-0.4 mg/ml,the oxidized self-assembled chitosan film substrate had highest antibody loading capacity.And it had a 3.94 fold higher antibody loading capacity than the ordinary aldehyde carrier.Conclusion：The agarose nano-membrane carrier is an ideal method for surface modification of immobilized antibody molecules,which is more suitable for preparation of immune microarray carrier.

关 键 词：琼脂糖自组装膜载体 免疫芯片 抗体固定 荧光强度 

分 类 号：Q816[生物学—生物工程]