Fast determination of haptoglobin phenotype and calculation of hemoglobin binding capacity using high pressure gel permeation chromatography

Abstract

A new and fast method for haptoglobin (Hp) phenotyping was developed based on high pressure gel permeation chromatography of hemoglobin-supplemented serum. Haptoglobin phenotypes 1-1, 2-1, and 2-2 are resolved on the difference in size of their hemoglobin–haptoglobin complexes. Results are available in less than 15 min. Results of the chromatographic typing correspond to those obtained by conventional starch gel electrophoresis. Next to the phenotyping of haptoglobin, the method allows reproducible calculation of the hemoglobin binding capacity (HBC) of human serum. Using this methodology, reference values for HBC were found to be 0.75±0.25 g/l, with the lowest HBC in Hp 2-2 subjects and the highest in Hp 1-1 subjects ( P<0.05). In contrast to earlier findings, the ratio HBC:Hp concentration was found to be comparable for the three Hp types. In conclusion, this method allows a rapid phenotyping in critical clinical conditions where Hp phenotyping can be useful, e.g. determining the donor’s phenotype in liver transplantation.