Development of a polymer-enzyme immunoassay method and its application.
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.
Liu
F
F
Department of Chemistry, Wuhan University, People's Republic of China.
Liu
F H
FH
Zhuo
R X
RX
Peng
Y
Y
Deng
Y Z
YZ
Zeng
Y
Y
D000180:Acrylic Resins; D000818:Animals; D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D005069:Evaluation Studies as Topic; D006514:Hepatitis B Surface Antigens / Q000097:blood; D006801:Humans; D018796:Immunoconjugates; D007124:Immunoenzyme Techniques / Q000706:statistics & numerical data; D008567:Membranes, Artificial; D051379:Mice; D012680:Sensitivity and Specificity; D013696:Temperature
D000180:Acrylic Resins; D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D006514:Hepatitis B Surface Antigens; D018796:Immunoconjugates; D008567:Membranes, Artificial; C052970:poly-N-isopropylacrylamide