乙酰胆碱和胆碱化学发光生物传感器的研究

A Chemoluminescence Biosensor for Determination of Acetylcholine and Choline

将具有分子识别功能的乙酰胆碱酯酶和胆碱氧化酶及进行换能反应的luminol和Cu^(2+)分别固定在多孔玻璃和离子交换剂的柱中,组成流动注射式胆碱和乙酰胆碱化学发光传感器,让传感器分子识别反应和换能反应在各自最佳pH值下进行.这一新型生物传感器优化了发光量子产率,避免了在传感元件上直接发光所产生的散射干扰.测定乙酰胆碱和胆碱的线性范围达到l～1000pmol,检测限为500fmol,每次测定时间为2min,寿命为6个月,已用于鼠脑及人血清中乙酰胆碱和胆碱的测定.

A chemluminescence(CL) biosensor responding to acetylcholine(ACh) and choline(Ch) was prepared by covalently coupling acetylcholinesterase(ACHE) and choline oxidase(ChO) to the controlled pore glass as a molecular recognition element, using electrostatically immobilizing luminol and copper ion exchange resin as a transduction element in a flow injection system. The analystes were injected into a continuous stream of simple medium flowing through a sequence of enzyme reactors containing AChE and ChO to produce hydrogen peroxide. The luminol and Cu + were eluted by Na2SO4 from the ion exchange resin column, then reacted with hydrogen peroxide to produce CL. The linear range for both the ACh and Ch was 1 ~ 1000 pmol, The detection limit was 500 fmol. A complete analysis can be finshed within 2 min. The sensor was stable for 6 months and has been applied successfully to the analysis of ACh and Ch in human serum and rat brain.