新型pH敏感相分离高分子的制备及其在免疫分析中的应用

聚Ｎ 异丙基丙烯酰胺 (ＰＮＩＰ)温度敏感高分子以其独特的温度敏感性质已成功地应用于免疫分析[1～ 6] .但这类温度敏感相分离免疫分析的反应温度必须控制在ＰＮＩＰ的相转变温度 ( 31℃ )以下 ,这不可避免地会影响免疫反应速率 .ｐＨ敏感高分子是另一类对环境敏感的智能型高分子 ,在其相转变ｐＨ值附近发生沉淀与溶解的可逆性变化 .目前 ,ｐＨ敏感高分子在免疫分析中的应用并没有受到重视 ,研究得较少[7] .这主要是由于 ｐＨ敏感高分子的相转变 ｐＨ值大都在 3左右 ,在此ｐＨ条件下 ,免疫反应生成的抗原 抗体免疫复合物会受到不同程度的…

Preparation of Novel pH Sensitive Phase Separating Polymer and Its Application in Fluorescence Immunoassay

Poly (N-isopropylacrylamide-methylacrylic acid) [P (NIP-co-MAA) ], a linear water-soluble pH precipitating synthetic polymer, was used as a novel separating carrier for the reactants in immunoassay.The P(NIP-co-MAA) precipitates out of water below a critical pH 5.6 at 37 ℃, and redissolved when the solution pH is above 6. 1. This characteristic of the P(NIP-co-MAA) made it possible to carry out the immunochemical steps of the assay in true solution and then to quickly separate the resulting procucts from the reaction mixtures. The above approach was applied to an assay for the sandwich immunoassay of hepatitis B surface antigen(HBsAg). Sensitivity of this method was close to that of traditional ELISA using same reactants. However, the assay was much faster(assay time decreased from 100－120 min to 45 min). This method has been applied to the determination of the HBsAg levels in human blood serum with satisfactory results. This general technique may also be used for a wide variety of separation processes in addition to immunoassay, in which a specific component is to be isolated for analysis, recovery, or disposal.