Graduate School of Science and Engineering (Science Section) and Venture Business Laboratory, Ehime University, Matsuyama, Japan.
J Pharm Biomed Anal. 2011 Dec 15;56(5):1085-8. doi: 10.1016/j.jpba.2011.07.027. Epub 2011 Jul 29.
Haptoglobin is known to bind to hemoglobin during intravascular hemolysis. Membrane-immobilized anti-haptoglobin antibody, which was produced after antibody was isolated by non-denaturing two-dimensional electrophoresis, was transferred to a polyvinylidene difluoride membrane and was stained using Ponceau S. The proteins bound to the membrane-immobilized anti-haptoglobin antibody were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and matrix-assisted laser desorption ionization/time-of-flight mass spectrometry. Hemoglobin was specifically obtained when the membrane-immobilized anti-haptoglobin antibody was incubated with human serum obtained from hemolysis blood. Furthermore, hemoglobin in the flowing fluid was captured by the membrane-immobilized anti-haptoglobin antibody and analyzed directly. The results indicate that hemolysis can be examined by direct trapping and analysis of hemoglobin under physiological conditions.
触珠蛋白在血管内溶血时与血红蛋白结合。非变性二维电泳分离抗体后，制备的膜固定抗触珠蛋白抗体被转移到聚偏二氟乙烯膜上，并用丽春红 S 染色。膜固定抗触珠蛋白抗体结合的蛋白质通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和基质辅助激光解吸电离/飞行时间质谱进行分析。当膜固定抗触珠蛋白抗体与溶血血液中获得的人血清孵育时，特异性获得血红蛋白。此外，流动液中的血红蛋白被膜固定抗触珠蛋白抗体捕获并直接分析。结果表明，在生理条件下可以通过直接捕获和分析血红蛋白来检查溶血。
