Aslan Kadir, Geddes Chris D
Institute of Fluorescence, Laboratory for Advanced Medical Plasmonics, Medical Biotechnology Center, University of Maryland Biotechnology Institute, 725 West Lombard Street, Baltimore, Maryland 21201, USA.
Anal Chem. 2007 Mar 1;79(5):2131-6. doi: 10.1021/ac0620967. Epub 2007 Jan 26.
In this paper, the proof of principle of microwave-accelerated aggregation assay technology, which shortens the solution-based aggregation assays' run time to seconds (>100-fold increase in kinetics) with microwave heating, was demonstrated using a model aggregation assay based on the well-known interactions of biotin and avidin. Biotinylated gold colloids were aggregated in solution with the addition of streptavidin, which takes 20 min at room temperature to reach >90% completion and only 10 s with microwave heating. The initial velocity (after 1-s microwave heating) of the biotinylated gold colloids reaches up to 10.5 m/s, which gives rise to greater sampling of the total volume but not a large increase in bulk temperature. The room-temperature, steady-state velocity of the colloids was <0.5 microm/s. In control experiments, where streptavidin preincubated with d-biotin in solution is added to biotinylated gold colloids and microwave heated, gold colloids did not aggregate, demonstrating that nonspecific interactions between biotinylated gold colloids and streptavidin were negligible.
在本文中,基于生物素和抗生物素蛋白的著名相互作用,通过一个模型聚集试验,证明了微波加速聚集试验技术的原理验证。该技术利用微波加热将基于溶液的聚集试验运行时间缩短至数秒(动力学提高超100倍)。在添加链霉亲和素的情况下,生物素化金胶体在溶液中聚集,在室温下这需要20分钟才能达到>90%的完成度,而微波加热仅需10秒。生物素化金胶体的初始速度(微波加热1秒后)高达10.5米/秒,这使得能够对总体积进行更多采样,但不会使整体温度大幅升高。胶体在室温下的稳态速度<0.5微米/秒。在对照实验中,将在溶液中与d -生物素预孵育的链霉亲和素添加到生物素化金胶体中并进行微波加热,金胶体未发生聚集,这表明生物素化金胶体与链霉亲和素之间的非特异性相互作用可忽略不计。