title: Direct trapping and analysis of hemoglobin in flowing fluid using membrane-immobilized anti-haptoglobin antibody.
abstract: 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.
authors.affiliation: Graduate School of Science and Engineering (Science Section) and Venture Business Laboratory, Ehime University, Matsuyama, Japan. shimazaki.yoji.my@ehime-u.ac.jp
authors.forename: Youji
authors.initials: Y
authors.lastname: Shimazaki
authors.forename: Ayaka
authors.initials: A
authors.lastname: Kimura
mesh_terms: D004591:Electrophoresis, Polyacrylamide Gel; D006242:Haptoglobins / Q000276:immunology*; D006454:Hemoglobins / Q000032:analysis*; D006801:Humans; D008567:Membranes, Artificial; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D006242:Haptoglobins; D006454:Hemoglobins; D008567:Membranes, Artificial