Takátsy Anikó, Sedzik Jan, Kilár Ferenc, Hjertén Stellan
Institute of Bioanalysis, Faculty of Medicine, University of Pécs, Szigeti út, Pécs, Hungary.
J Sep Sci. 2006 Dec;29(18):2810-5. doi: 10.1002/jssc.200600212.
Artificial and highly selective antibodies (in the form of gel granules) against proteins can easily be synthesized by a simple, cost-effective imprinting technique [Liao, J.-L. et al., Chromatographia 1996, 42, 259-262]. Using the same method for synthesis of gel antibodies against viruses in combination with analysis by free zone electrophoresis in a rotating narrow bore tube we have shown that artificial gel antibodies against Semliki Forest Virus (wild type) can sense the difference between this virus and a mutant, although they differ in their chemical composition only by three amino acids in one of the three proteins on the surface of the virus particle. The reason for this extremely high resolution is explained by the fact that we use three types of selectivity: (i) shape selectivity (created by the close fit between the antigen and its imprint in the gel), (ii) bond selectivity in the contact area between the antigen and its imprint in the gel antibody, and (iii) charge selectivity, originating from slightly different structures or/and conformations of the antigens.
通过一种简单且经济高效的印迹技术,能够轻松合成针对蛋白质的人工且高度选择性的抗体(呈凝胶颗粒形式)[廖,J.-L. 等人,《色谱学》1996 年,42 卷,259 - 262 页]。运用相同方法合成针对病毒的凝胶抗体,并结合在旋转窄孔管中进行的自由区电泳分析,我们已表明,针对塞姆利基森林病毒(野生型)的人工凝胶抗体能够识别该病毒与其突变体之间的差异,尽管它们在化学组成上仅在病毒颗粒表面三种蛋白质之一中相差三个氨基酸。这种极高分辨率的原因可由以下事实解释:我们使用了三种类型的选择性:(i)形状选择性(由抗原与其在凝胶中的印迹紧密契合产生),(ii)抗原与其在凝胶抗体中的印迹之间接触区域的键合选择性,以及(iii)电荷选择性,其源于抗原结构或/和构象的微小差异。