A new method for fast haptoglobin phenotyping and hemoglobin binding capacity calculation based on capillary zone electrophoresis.
A capillary zone electrophoresis method was developed for haptoglobin (Hp) phenotyping in hemoglobin (Hb) supplemented serum. The method allows a complete resolution of the major haptoglobin phenotypes Hp 1-1, Hp 2-1, and Hp 2-2 based on the difference in charge-to-mass ratio of their Hb-Hp complexes. Identification of these phenotypes was achieved by their significant differences in migration times and their marked difference in electrophoretic pattern. Our method showed full agreement with starch gel electrophoresis. Furthermore, following neuraminidase treatment of the serum, the Hp subtypes Hp1-1FF, FS and SS could be resolved, based on the same criteria as the phenotyping. The new electrophoretic method allowed typing of the rare phenotypes Hp 2-1 modified (Hp 2-1M) and Hp Johnson. The calculated hemoglobin binding capacity of serum correlates well with the nephelometrically determined haptoglobin concentration. The new method for typing haptoglobin gives prospectives for fast haptoglobin typing and Hp 1-1 subtyping.
Wuyts
B
B
Department of Clinical Chemistry, Gent University Hospital, Belgium.
Delanghe
J R
JR
Kasvosve
I
I
Langlois
M R
MR
De Buyzere
M L
ML
Janssens
J
J
D019075:Electrophoresis, Capillary / Q000379:methods; D004592:Electrophoresis, Starch Gel; D006242:Haptoglobins / Q000737:chemistry / Q000145:classification* / Q000235:genetics / Q000378:metabolism*; D006454:Hemoglobins / Q000737:chemistry / Q000145:classification* / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007525:Isoelectric Focusing; D009391:Nephelometry and Turbidimetry / Q000379:methods; D009439:Neuraminidase / Q000378:metabolism; D010641:Phenotype; D011110:Polymorphism, Genetic; D011485:Protein Binding; D012680:Sensitivity and Specificity
D006242:Haptoglobins; D006454:Hemoglobins; C035492:haptoglobin-hemoglobin complex; D009439:Neuraminidase