Abelev G I, Karamova E R
Anal Biochem. 1984 Nov 1;142(2):437-44. doi: 10.1016/0003-2697(84)90487-1.
Discontinuous electrophoresis on cellulose acetate membranes with the use of 0.06 M Tris-HCl (pH 6.7) as the leading electrolyte and 0.012 M Tris-beta-alanine (pH 8.6) as the terminating one results in concentration of the proteins present in the system on the Cl-/beta-alanine- boundary. If the antigen solution is placed in a "pocket" ahead of the moving boundary, a counterflow to the cathode arises due to electroendosmosis. At constant voltage the migration rate of the boundary drops and that of electroendosmosis does not change until they become equal. In such a stationary position, the antigen-containing solution is passing through the Cl-/beta-alanine- boundary to the cathode, while all the proteins are completely "absorbed" on the boundary as highly concentrated bands. Addition of ampholytes to the antigen solution contributes to the isotachophoretic separation of a protein mixture on the strip. The concentrated and separated antigens can be revealed by immunofixation, immunodiffusion, or crossed immunoelectrophoresis in gel. The technique is approximately 100 times more sensitive compared to the usual immunodiffusion and immunoelectrophoresis methods on cellulose acetate membranes, and is applicable to the detection of trace amounts of antigens in the urine, liquor, amniotic fluid, tears, and other biological fluids with low protein contents.
在醋酸纤维素膜上进行间断电泳，使用0.06 M Tris-HCl（pH 6.7）作为前导电解质，0.012 M Tris-β-丙氨酸（pH 8.6）作为终止电解质，会使系统中存在的蛋白质在Cl⁻/β-丙氨酸边界处浓缩。如果将抗原溶液置于移动边界前方的“口袋”中，由于电渗作用会产生向阴极的逆流。在恒定电压下，边界的迁移速率下降，而电渗速率直到二者相等时才会改变。在这样的静止位置，含抗原的溶液通过Cl⁻/β-丙氨酸边界流向阴极，而所有蛋白质都作为高度浓缩的条带完全“吸附”在边界上。向抗原溶液中添加两性电解质有助于在条带上对等速电泳分离蛋白质混合物。浓缩和分离后的抗原可通过免疫固定、免疫扩散或凝胶中的交叉免疫电泳来揭示。与在醋酸纤维素膜上进行的常规免疫扩散和免疫电泳方法相比，该技术的灵敏度大约高100倍，适用于检测尿液、脑脊液、羊水、眼泪及其他蛋白质含量低的生物体液中的痕量抗原。
