Ullman E F, Kirakossian H, Singh S, Wu Z P, Irvin B R, Pease J S, Switchenko A C, Irvine J D, Dafforn A, Skold C N
Research Department, Syva Company, Palo Alto, CA 94303.
Proc Natl Acad Sci U S A. 1994 Jun 7;91(12):5426-30. doi: 10.1073/pnas.91.12.5426.
A method for monitoring formation of latex particle pairs by chemiluminescence is described. Molecular oxygen is excited by a photosensitizer and an antenna dye that are dissolved in one of the particles. 1 delta gO2 diffuses to the second particle and initiates a high quantum yield chemiluminescent reaction of an olefin that is dissolved in it. The efficiency of 1 delta gO2 transfer between particles is approximately 3.5%. The technique permits real-time measurement of particle binding kinetics. Second-order rate constants increase with the number of receptor binding sites on the particles and approach diffusion control. By using antibody-coated particles, a homogeneous immunoassay capable of detecting approximately 4 amol of thyroid-stimulating hormone in 12 min was demonstrated. Single molecules of analyte produce particle heterodimers that are detected even when no larger aggregates are formed.
描述了一种通过化学发光监测乳胶颗粒对形成的方法。分子氧由溶解在其中一个颗粒中的光敏剂和天线染料激发。单线态氧扩散到第二个颗粒并引发溶解在其中的烯烃的高量子产率化学发光反应。颗粒之间单线态氧转移的效率约为3.5%。该技术允许实时测量颗粒结合动力学。二级速率常数随颗粒上受体结合位点的数量增加而增加，并接近扩散控制。通过使用抗体包被的颗粒，证明了一种能够在12分钟内检测约4阿摩尔促甲状腺激素的均相免疫测定法。即使没有形成更大的聚集体，分析物的单个分子也会产生可检测的颗粒异二聚体。
