Analysis of hemoglobin and globin chain variants by a commonly used capillary isoelectric focusing method.

摘要: To analyze both hemoglobin (Hb) and globin chain variants, we modified a commonly used method, capillary isoelectric focusing (CIEF), with detection at 280 nm. The samples were hemolysates prepared from red blood cells, chains obtained the by treatment cold acidified acetone. When migration time for internal reference, carbonic anhydrase I (isoelectric point, pI 6.60), was taken as 1.0, ratio Hb A0 in normal human 0.877 +/- 0.004 (mean SD, n = 9), those of alpha- beta-globin 0.673 0.847 0.005 4), respectively. peak heights between beta- alpha-globin (beta/alpha) Hbs four subjects almost constant 2.5 0.1 SD). This indicates which includes mutation (if one exists). an variant, Hoshida (in Gln is substituted Glu residue 43 chain), analyzed this two main peaks observed (migration ratios 0.836 0.877, corresponding to abnormal Hb, respectively). An additional 0.788 also detected profiles. 1.57, indicating that exists chain. We thus established convenient system using CIEF provides rapid reproducible method random analysis variants.