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Development of a polymer‐enzyme immunoassay method and its application
Author(s) -
Liu F.,
FH Liu,
RX Zhuo,
Peng Y.,
YZ Deng,
Zeng Y.
Publication year - 1995
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1111/j.1470-8744.1995.tb00335.x
Subject(s) - immunoassay , chemistry , horseradish peroxidase , chromatography , antibody , enzyme , homogeneous , peroxidase , antigen , conjugate , hbsag , reagent , biochemistry , hepatitis b virus , immunology , organic chemistry , biology , mathematical analysis , physics , virus , mathematics , thermodynamics
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.