新型均相电化学发光免疫分析法研究

The immunoassay is an front research subject of modern analytical chemistry that has been widely employed in life science and in environment science due to high specificity and high sensitivity. Electrogenerated chemiluminescence (ECL) immunoassay (ECLIA) is a promising analytical tool in immunossay. The project, "novel homogeneous ECL immunoassays", has focused on three research sections including the investigation of new ECL systems, electron transfer mediators and the development of homogeneous ECL immmunoassy. Two ECL methods of the determination of riboflavin and adrenson have been developed based on the ECL of lucigenin and Mn(Ⅲ), respectively. A novel flow injection analytical system with a screen-printed enzyme sensor incorporating Os-complex mediator was developed and a glucose amperometric sensor with an integrated pre-oxidation membrane to remove interference from ascorbic acid and uric acid was fabricated. Two novel homogeneous electrogenerated chemiluminescence (ECL) immunoassays for anti-digoxin antibody and digoxin hapten were developed based on a luminol-labeled digoxin hapten and a multilabeling of luminol and digoxin to a bovine serum albumin, respectively. Digoxin hapten was determined throughout its therapeutic range with a detection limit of 0.10 ng/ml and in a 3 orders of magnitude range. The proposed methods hvae been applied to assay anti-digoxin antibody and digoxin in human control serum with satisfactory results. The project achievement includes 4 research papers published on SCI journals, 3 research papers on Chinese key journals and two presentations on 2002 Pittcon international conference and the goal of the project was mainly accomplished. The project achievements, homogeneous ECL immunoassay using luminuol-labeled small molecule hapten and multilabeling of luminol and digoxin, are promising alternative approach for improvement of the ECL quantum, ECL label ratio and the sensitivity of ECL immunoassay. The proposed ECL immunoassay for digoxin and anti-digoxin antibody is easily performed with high sensitivity and has a potential use in clinical assay and will promote the research work on the immunoassay in some aspects.

提出基于小分子氧化还原标记偶合化学发光检测的新型均相电化学催化化学发光免疫分析法。筛选新型小分子电催化化学发光免疫标记物，研究标记蛋白免疫反应特性和电化学发光特性，建立新型均相电化学催化化学发光免疫分析理论模式和快速简单高灵敏自动化均相电催化化学发光免疫分析方法。对生命科学和环境科学研究以及免疫分析的发展均具有重要的意义。