Pollard-Knight D, Hawkins E, Yeung D, Pashby D P, Simpson M, McDougall A, Buckle P, Charles S A
Amersham International, Pollards Wood Laboratories, Bucks, UK.
Ann Biol Clin (Paris). 1990;48(9):642-6.
A technique based on surface plasmon resonance is described which can be used to detect changes of refractive index that occur when one partner of a molecular binding pair diffuses from solution to bind the other partner which is immobilised on a silver surface. Results for the molecular binding pairs; protein-antibody, hapten-antibody and DNA-DNA are described. Instrumentation necessary for implementation of the technique is detailed. Immunoassay of proteins and haptens is possible in less than one minute with a sensitivity of 10(-9) mol/l. Hybridisation of 10 fmoles of a 97 base target sequence on the 1 mm2 area of detection to an immobilised oligonucleotide probe can be detected in less than five minutes. Advantages of the technique include the ability to record the kinetics of binding reactions in "real time" and the lack of labels in this simple assay format. Methods of improving the sensitivity are discussed.
本文描述了一种基于表面等离子体共振的技术,该技术可用于检测当分子结合对中的一个组分从溶液中扩散出来与固定在银表面的另一个组分结合时发生的折射率变化。文中描述了蛋白质-抗体、半抗原-抗体和DNA-DNA等分子结合对的检测结果。详细介绍了实施该技术所需的仪器设备。蛋白质和半抗原的免疫测定可在不到一分钟内完成,灵敏度为10^(-9)摩尔/升。在不到五分钟的时间内,可以检测到10飞摩尔的97个碱基的靶序列在1平方毫米的检测区域上与固定化寡核苷酸探针的杂交。该技术的优点包括能够“实时”记录结合反应的动力学,以及这种简单检测形式中无需标记。文中还讨论了提高灵敏度的方法。