琼脂糖凝胶蛋白芯片片基的制备及应用  

Preparation of Agarose Film-Barotein Microarray and Its Application in Detection for Hepatitis B Virus Antigens

作　　者：刘志红[1] 何为[1] 韩旭[1] 许丹科[1] 

机构地区：[1]军事医学科学院放射医学研究所,北京100850

出　　处：《分析化学》2007年第5期775-778,共4页Chinese Journal of Analytical Chemistry

基　　金：国家科技攻关计划项目"人类重大疾病及重要生理功能相关的蛋白质组学研究"资助(2002-BA711A11)

摘　　要：建立了一种制备琼脂糖凝胶修饰玻片的方法。以乙肝表面抗原为检测指标,比较了琼脂糖凝胶片基和常规的醛基片基固定抗体后的检测灵敏度,发现琼脂糖凝胶修饰后可以明显增加检测的荧光强度,并在10ng/L～100μg/L之间具有良好的线性关系。在此片基同时固定了抗HBsAg和抗HBeAg抗体,并以牛血清白蛋白和生物素标记抗体为阴性和阳性对照点,采用夹心式免疫反应法检测乙肝病毒的两种抗原,取得良好效果。A novel type of modified layer based on agarose film was developed for fabrication of antibody microarray. The surface of glass slides was modified with agarose solution and the resulting various agarose films by using different concentration of agarose solution were analyzed by atomic force microscopy. The results showed that 1% agarose solution can produce the optimum performance for antibody microarray such as mechanical strength and micropore structure. As proof-of-principle experiments, the modified slide was further used to prepare anti-hepatitis B virus surface antigen antibody microarrays. A detection limit of 10 ng/L and a calibration concentration range from 10 ng/L to 100 μg/L could be obtained for hepatitis B virus surface antigen. In addition, this antibody microarray was further compared to the one based on an aldehyde modified layer. As a result, the agarose film could increase ted with anti-HBsAg（ anti-hepatitis B virus surface the detection sensitivity significantly. The microarray spotantigen）, anti HBeAg （anti-hepatitis B virus e antigen）, negtive control and positive control was assayed to HBsAg , HBeAg and the mixture, respectively. The resultsshowed a satisfied discrimination.

关 键 词：琼脂糖凝胶薄膜 蛋白微阵列芯片 乙肝病毒抗原检测 

分 类 号：Q819[生物学—生物工程]