New Direction for Enzyme-Linked Binding/Immunoassays

该项目由分析和表面化学项目支持，旨在开发用于生物大分子选择性定量的酶免疫测定型传感器。在这项为期三年的标准资助期间，Meyerhoff教授和他在密歇根大学的学生将研究硫辛酸自组装单层结合酶抗体电极的操作基础，该电极能够分析未经处理的全血中的蛋白质抗原。介绍了具有在未稀释全血中以万亿分之一的水平测定人绒毛膜促性腺激素（hCG）和前列腺特异性抗原（PSA）的能力的新型传感器。未来的努力将面向传感器设备的小型化，这些设备能够在广泛的动态浓度范围内对蛋白质分析物进行自动化的多组分分析。此外，Meyerhoff教授还继续致力于开发酶联结合测定法（ELBAs），通过这种方法可以表征与糖胺聚糖特异性相互作用的带电生物大分子的络合和结合位点。后一种技术用于碳水化合物结构和完整糖蛋白含量的评估。很少有分析探针能与酶抗体型传感器对生物大分子的敏感性和选择性相媲美。这些设备能够检测存在于复杂基质（如全血和废水）中的蛋白质分析物。Meyerhoff教授和他在密歇根大学的学生正在开发新一代的仪器，这些仪器能够在较宽的浓度范围内用最少的样品预处理分析蛋白质分析物的混合物。该分析技术将广泛应用于生物技术、临床化学和药物研究和生产。

This project, supported in the Analytical and Surface Chemistry Program, is directed towards the development of enzyme immunoassay type sensors for the selective quantification of biomacromolecules. During the tenure of this three-year standard grant, Professor Meyerhoff and his students at the University of Michigan will investigate the fundamentals of operation of thioctic acid self-assembled monolayer bound enzyme-antibody electrodes capable of assaying protein antigens in untreated whole blood. Novel sensors are introduced having the capability of assaying human chorionic gonadotropin (hCG) and prostate specific antigen (PSA) at the part-per-trillion level in undiluted whole blood. Future efforts will be geared towards the miniaturization of sensor devices which are capable of automated, multi-component analyses of protein analyte over a wide dynamic concentration range. In addition, Professor Meyerhoff is also continuing his efforts in developing enzyme- linked binding assays (ELBAs) whereby the complexation and binding sites of charged biological macromolecules that interact specifically with glycosaminoglycans can be characterized. The latter techniques are used in the assessment of carbohydrate structure and content of intact glycoproteins. Few analytical probes can rival the sensitivity and selectivity for biological macromolecules as exhibited by enzyme-antibody type sensors. These devices are capable of sub-part-per-trillion detection of protein analyte present in complex matrices such as whole blood and waste water. Professor Meyerhoff and his students at the University of Michigan are advancing new generations of these devices which are capable of assaying mixtures of protein analytes over wide concentration ranges with minimal sample pretreatment. This analytical technology will be broadly applicable to biotechnology, clinical chemistry, and pharmaceutical research and manufacture.