Kim K Y, Han M Y, Yoon D Y, Cho B Y, Choi M J, Choe I S, Chung T W
Laboratory of Cell Biology, Genetic Engineering Research Institute, Korea Institute of Science and Technology, Daejon.
Immunol Lett. 1992 Feb 15;31(3):267-72. doi: 10.1016/0165-2478(92)90125-8.
A fluoroimmunoassay employing a flow cytometer as the fluorescence-detecting device is described. Three kinds of antigens, murine immunoglobulin (Ig), human chorionic gonadotropin (hCG) and progesterone were chosen as examples of the assay using fluorescein-labeled antibodies. Cyanogen bromide-activated agarose beads were used as solid-phase supporters. The flow cytometric immunoassay was applied to both qualitative and quantitative analyses; determination of murine Ig isotypes, quantitative determination of Ig, hCG and a hapten, progesterone. This assay produced very reproducible and less-fluctuating data since thousands of particles in the assay were collected and processed to produce a single value for fluorescence intensities. Furthermore, the working range of the assay in terms of antigen concentration was much broader than that of enzyme immunoassay. Therefore, we believe that microparticles like agarose beads could be useful solid-phase supporters in immunoassay, and the flow cytometer could provide a reliable alternative to the fluorescence-detecting device in immunoassay.
描述了一种使用流式细胞仪作为荧光检测装置的荧光免疫测定法。选择了三种抗原，即鼠免疫球蛋白（Ig）、人绒毛膜促性腺激素（hCG）和孕酮，作为使用荧光素标记抗体进行测定的示例。溴化氰活化的琼脂糖珠用作固相支持物。流式细胞免疫测定法用于定性和定量分析；测定鼠Ig同种型、定量测定Ig、hCG和一种半抗原孕酮。由于在测定中收集并处理数千个颗粒以产生单个荧光强度值，该测定产生了非常可重复且波动较小的数据。此外，该测定法在抗原浓度方面的工作范围比酶免疫测定法宽得多。因此，我们认为像琼脂糖珠这样的微粒可以成为免疫测定中有用的固相支持物，并且流式细胞仪可以为免疫测定中的荧光检测装置提供可靠的替代方案。
