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A novel fluorescence immunoassay system based on pH-sensitive phase separating technique

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A novel fluorescence immunoassay system based on pH-sensitive phase separating technique.htm (413bytes)
Date
2005-04
Author
Lin Peng
Zheng Hong
郑洪
Yang Huang-Hao
Li Dong-Hui
Xu Jin-Gou
许金钩
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  • 化学化工-已发表论文 [14469]
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Abstract
In this paper, it was discovered that a novel pH-sensitive copolymer of N-isopropylacrylamide (NIP) and N-(3-dimethylaminopropyl)methacrylamide (DMAPM) could be gotten by polymerization. The phase transition pH (pH(tr)) of P(NIP-DMAPM) polymer was found to be 7.4 at 37 degreesC. The polymer was precipitated out of water above a critical pH=7.4 and re-dissolved below pH=7.4. The characteristic of this polymer made it possible to carry out the immunochemical steps of an immunoassay in a true solution and then to quickly separate the resulting product from the reaction mixture. In a competitive fluorescence immunoassay, the standard rabbit IgG and rabbit IgG immobilized on P(NIP-DMAPM) first competitively reacted with the fluorescein isothiocyanate (FITC) labeled antibody, then the pH of solution was adjusted above the pHtr of polymer to precipitate the polymer-immune complex, and the polymer-immune complex precipitate was separated and re-dissolved by the adjustment of pH, finally the FITC-labeled antibody in the immune complex was quantified by fluorescence measurement. The calibration graph for rabbit IgG was linear over the range of 100-1000 ng/mL with a detection limit of 11 ng/mL. The method is rapid, sensitive and simple. Owing to neutral pH(tr) of P(NIP-DMAPM), the damage to antigen-antibody immune complex was greatly decreased in the course of separation. In addition, a sandwich enzyme-linked fluorescence immunoassay method for the determination of human IgG was also developed, showing that the pH-sensitive phase separating immunoassay could be performed in the competitive method as well as the sandwich method.
Citation
CHINESE JOURNAL OF CHEMISTRY,2005,23(3):285-290
URI
http://dx.doi.org/doi:10.1002/cjoc.200590285
https://dspace.xmu.edu.cn/handle/2288/11249

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