Liu F, Liu F H, Zhuo R X, Peng Y, Deng Y Z, Zeng Y
Department of Chemistry, Wuhan University, People's Republic of China.
Biotechnol Appl Biochem. 1995 Jun;21(3):257-64.
Both poly(N-isopropylacrylamide) and poly(N-isopropylacrylamide)-antibody (PINP-Ab)-labelled enzyme adhered quickly and tightly to cellulose acetate/nitrate membrane either below (less efficiently) or above (more efficiently) the lower critical solution temperature, and the retention of PINP-Ab on the membrane increased over 30-fold when compared with the unconjugated Ab. These characteristics were used to develop a novel polymer-enzyme-linked immunoassay method: homogeneous antigen-antibody immune-complexation reaction and a heterogeneous separation process. By using a simple horseradish-peroxidase-labelled antibody as a probe, we applied this method to the detection of human serum hepatitis B surface antigen (HBsAg). This immunoassay system can detect as little as 1 ng/ml of HBsAg. The advantages of this method are: (a) fast homogeneous immune complexation; (b) a rapid heterogeneous separation process; (c) high sensitivity; and (d) low non-specific background.
聚（N-异丙基丙烯酰胺）和聚（N-异丙基丙烯酰胺）-抗体（PINP-Ab）标记的酶在低于（效率较低）或高于（效率较高）低临界溶液温度时，都能快速且紧密地附着在醋酸纤维素/硝酸纤维素膜上，并且与未偶联的抗体相比，PINP-Ab在膜上的保留量增加了30多倍。利用这些特性开发了一种新型的聚合物酶联免疫分析方法：均相抗原-抗体免疫复合反应和异相分离过程。通过使用简单的辣根过氧化物酶标记抗体作为探针，我们将该方法应用于检测人血清乙肝表面抗原（HBsAg）。这种免疫分析系统能够检测低至1 ng/ml的HBsAg。该方法的优点包括：（a）快速的均相免疫复合；（b）快速的异相分离过程；（c）高灵敏度；（d）低非特异性背景。
