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

A new and fast method for haptoglobin (Hp) phenotyping was developed based on high pressure gel permeation chromatography of hemoglobin-supplemented s erum. Haptoglobin phenotypes 1-1, 2-1, and 2-2 are resolved on the differen ce in size of their hemoglobin-haptoglobin complexes. Results are available in less than 15 min. Results of the chromatographic typing correspond to t hose obtained by conventional starch gel electrophoresis. Next to the pheno typing of haptoglobin, the method allows reproducible calculation of the he moglobin binding capacity (HBC) of human serum. Using this methodology, ref erence 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 contr ast to earlier findings, the ratio HBC:Hp concentration was found to be com parable for the three Hp types. In conclusion, this method: allows a rapid phenotyping in critical clinical conditions where Hp phenotyping can be use ful, e.g. determining the donor's phenotype in liver transplantation. (C) 2 000 Elsevier Science B.V. All rights reserved.