Comparison of Immobilization Modes in pH-Sensitive Phase Separation Immunoassay
Three immobilization modes of antigen to the polymers in the pH-sensitive phase separation immunoassay were investigated and compared. The results showed that the immobilization mode in the presence of N-ethyl-N'(3-dimethylaminopropyi)carbodiimide hydrochloride (EDCI) rendered the most desirable results. The immobilization efficiencies and immunological reaction activities of immobilized antigen of this mode were improved over the other two modes. The novel immobilization mode by EDCI was used in the pH-sensitive phase separation immunoassay for rabbit IgG (Ag). In the competitive immunoassay, immobilized Ag and the standard Ag (or sample) competed for binding to a horseradish peroxidase labeled antibody at 37 ℃ in a homogeneous format.Afar changing the pH to separate the polymer-immune complex, the complex precipitate was re-dissolved and determined by coupling with the color reaction of hydrogen peroxide and o-phenylenediamine. The linear range of this determination was between 100--1400 ng/mL. Compared to the traditional enzyme-linked immunosorbent assays (ELISA) using the same reactants, the proposed method was quiet fast (the time decreased from 100--120 to 30 min) and showed similar sensitivity, i.e., 6.0 ng/mL.
作 者： LIN, Peng GUO, Songlin WANG, Yilei WANG, Weigang CHEN, Jinmin JIA, Xiwei WANG, Guodong 作者单位： Key Laboratory of Science and Technology for Aquaculture and Food Safety, Fisheries College, Jimei University, Xiamen, Fujian 361021, China 刊 名： 中国化学（英文版） SCI 英文刊名： CHINESE JOURNAL OF CHEMISTRY 年，卷(期)： 2009 27(11) 分类号： O6 关键词： pH-sensitive phase separating polymer immnnoassay rabbit IgG N-ethyl-N'-(3-dimethylaminopropyl)carbodiimide hydrochioride N-acryloxysuccinimide