Binding of biotinylated DNA to streptavidin-coated polystyrene latex: Effects of chain length and particle size
Binding of biotinylated DNA to streptavidin-coated polystyrene latex: Effects of chain length and particle size
复制标题
DOI：
10.1006/abio.1996.0208
复制
发表时间：
1996-05-15
影响因子：
2.9
通讯作者：
Caldwell, KD
中科院分区：
文献类型：
--
作者：
Huang, SC;Stump, MD;Caldwell, KD
用直径为0.944 - 0.090 μ m的链霉亲和素包被的聚苯乙烯乳胶颗粒研究了大小为100 - 5000碱基对的5 ′-末端生物素化DNA的结合。该固相反应的实验结合常数和正向速率常数被确定为比预期的生物素-链霉亲和素在溶液中的相互作用的值低几个数量级，并且被证明取决于配体和底物的大小。在生物素化的DNA的结合常数中观察到的拐点出现在1000个碱基对左右，这可能表明高于和低于这个临界尺寸的大配体的不同表面取向。对于本研究中使用的较小乳胶颗粒，这种效果更为明显，并突出了不同尺寸颗粒上链霉亲和素表面排列的可能差异。扩散限制的结合反应被认为是显着的，在所有情况下。在目前的工作中，实验结合常数和生物素化的DNA中的碱基对的数量之间建立了指数关系。这种关系可能提供了一种手段来预测的情况下，吸附，纯化和释放较大的DNA链的能力和结合速度。(C)出版社：Academic Press，Inc.
The binding of 5'-end biotinylated DNA, ranging in size hom 100 to 5000 base pairs, was studied using streptavidin-coated polystyrene latex particles with diameters between 0.944 and 0.090 mu m. The experimental binding constants and forward rate constants of this solid-phase reaction were determined to be several orders of magnitude lower than values for the biotin-streptavidin interaction in solution as expected and were shown to depend on the size of both Ligand and substrate. An observed inflection in the binding constant of the biotinylated DNA appeared around 1000 base pairs, possibly indicating different surface orientations of the macroligand above and below this critical size. This effect was more pronounced for the smaller latex particles used in this study and highlighted possible differences in the surface arrangement of streptavidin on the differently sized particles. Diffusion limitation to the binding reaction was found to be significant in all cases. In this present work, an exponential relationship was established between the experimental binding constant and the number of base pairs in the biotinylated DNA This relationship possibly provides a means to predict capacity and binding speed in cases where adsorption, purification, and release of larger DNA chains are required. (C) 1996 Academic Press, Inc.