New Direction for Enzyme-Linked Binding/Immunoassays

该项目得到了分析和表面化学计划的支持，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拨款期间，密歇根大学的迈耶霍夫教授和他的学生将研究硫辛酸自组装单层结合酶抗体电极的基本操作原理，该电极能够检测未经处理的全血中的蛋白质抗原。介绍了一种新型传感器，该传感器能够在未稀释的全血中检测人绒毛膜促性腺激素(HCG)和前列腺特异性抗原(PSA)。未来的努力将着眼于传感器设备的小型化，这种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、多组分分析。此外，迈耶霍夫教授还在继续开发酶连接结合分析(ELBAS)，利用该方法可以表征与糖胺聚糖特异相互作用的带电生物大分子的络合和结合部位。后一种技术用于评估碳水化合物结构和完整糖蛋白的含量。很少有分析探针能与酶-抗体类型传感器对生物大分子的灵敏度和选择性相媲美。这些设备能够对全血和废水等复杂基质中存在的蛋白质分析物进行万亿分之一以下的检测。密歇根大学的迈耶霍夫教授和他的学生正在开发新一代的这种设备，这种设备能够在最小的样品前处理情况下，在广泛的浓度范围内分析蛋白质分析物的混合物。这项分析技术将广泛应用于生物技术、临床化学和药物研究与制造。

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.