Determination of intermediates of hemoglobin-haptoglobin complex of haptoglobin polymers by "Crossed hemoglobin electrophoresis".
We designed a "crossed hemoglobin electrophoresis" to be able to detect the intermediates of a hemoglobin-haptoglobin complex without the need for the purification step of haptoglobin molecules. Native polyacrylamide gel electrophoresis was performed as the first dimensional electrophoresis followed by a cross to hemoglobin as the second dimensional electrophoresis. Using this method, we found that a number of intermediates of normal Hp 2-2, Hp 2-1 polymers, particularly those from the trimer to the pentamer, were detected on a polyacrylamide gel. Each of their polymers combined to the hexamer with at least an equimolar content of hemoglobin. In the process of detecting the haptoglobin serotype of more than 10,000 patients for the clinical utilization of quantitative analytical data, we observed two haptoglobin variants, the Hp Johnson and a haptoglobin serotype similar to the Carlberg type. Using our crossed hemoglobin electrophoresis, we also detected the capacity for combining with hemoglobin of each polymer of these variants.
Okazaki
T
T
Department of Forensic Science, School of Allied Health Sciences, Kitasato University, 1-15-1 Kitasato, Sagamihara City, 228, Japan.
Yanagisawa
Y
Y
Nagai
T
T
D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D005260:Female; D006242:Haptoglobins / Q000737:chemistry*; D006454:Hemoglobins / Q000737:chemistry*; D006801:Humans; D008297:Male; D010375:Pedigree
D006242:Haptoglobins; D006454:Hemoglobins; C035492:haptoglobin-hemoglobin complex