BIOSYST-MeBioS, Katholieke Universiteit Leuven, Willem de Croylaan 42, B-3001 Leuven, Belgium.
Molecules. 2010 Mar 2;15(3):1127-40. doi: 10.3390/molecules15031127.
We have selected aptamers binding to lysozyme from a DNA library using capillary electrophoresis-systematic evolution of ligands by exponential enrichment. During the selection process the dissociation constant of the ssDNA pool decreased from the micromolar to the low nanomolar range within five rounds of selection. The final aptamer had a dissociation constant of 2.8 +/- 0.3 nM, 6.1 +/- 0.5 nM, and 52.9 +/- 9.1 nM as determined by fluorescence anisotropy, surface plasmon resonance and affinity capillary electrophoresis respectively. The aptamers were successfully challenged for specificity against other egg white proteins. The high affinity aptamers open up possibilities for the development of aptamer based food and medical diagnostics.
我们从一个 DNA 文库中使用毛细管电泳-指数富集的配体系统进化（SELEX）方法筛选到了与溶菌酶结合的适体。在筛选过程中，ssDNA 库的离解常数在五轮筛选后从微摩尔降低到了纳摩尔级。最终的适体的离解常数通过荧光各向异性、表面等离子体共振和亲和毛细管电泳分别测定为 2.8 ± 0.3 nM、6.1 ± 0.5 nM 和 52.9 ± 9.1 nM。这些适体在针对其他蛋清蛋白的特异性方面得到了成功的挑战。高亲和力的适体为基于适体的食品和医疗诊断的开发开辟了可能性。
