用温度敏感高分子为载体的激光光声免疫分析及其应用  

Photoacoustic Immunoassay Using Temperature Responsive Polymer As Carrier and Its Application

作　　者：周平[1] 邓延倬[1] 曾云鹗[1] 

机构地区：[1]武汉大学化学系,分析测试科学系武汉430072

出　　处：《分析化学》1997年第2期144-148,共5页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金

摘　　要：将单克隆抗乙肝表面抗原抗体与温度敏感高分子聚N- 丙基丙烯酸胺连接，在液 相中与抗原、酶标抗体反应，通过热沉淀实现与游离酶标抗体的异相分离。高分子载体的引 入能减少非特异性吸附，较常用固相载体聚苯乙烯酶标板更有利于激光光声技术高灵敏度优 势的发挥。同时，采用0．5％的十二烷基硫酸钠水溶液作显色终止剂，易与He－Ne激光波长 匹配，实现了对低值乙肝表面抗原阳性血清的检测。A novel ultrasensitive immunoassay method that uses poly (N-isopro- pylacrylamide) as carrier and laser-induced photoacoustic quantitation is proposed. Ultra- trace determination by laser spectroscopy in immunoassay has often plagued by nonspecific binding to the solid phase. This obstruction can be suppressed by introducing temperature responsive polymer, poly(N-isopropylacrylamide). A monoclonal antibody which has been conjugated to the polymer reacts with antigen and enzyme-labelled antibody below the critical solution temperature of the polymer. The polymer/immune complex sandwich can precipitate from the solution above the critical temperature. In this system, it is possible to minimize the opportunities for nonspecific binding and enhance reaction rate. Furthermore, when we use TMB as the substate of horseradish peroxidase and SDS and Na2SO3 as stop reagents, the color is stable for long time and can match the absorption wavelength of He-Ne laser. The method was employed to detect HBsAg in serum. A significant difference from the back- ground was obtained for 3. 5 × 10-11 g/mL. This value was almost one order of magnitude better than results obtained by ELISA.

关 键 词：激光光声 免疫分析 高分子载体 

分 类 号：R446.6[医药卫生—诊断学]