Abstract
The zeta (ζ) and various gamma (γ) chains of Hb Portland I have been separated and isolated from blood samples obtained from neonates with hydrops fetalis due to homozygous α-thalassemia. By using developers containing acetonitrile, methanol, and potassium phosphate and either an analytical (3.9 mm × 30 cm) or a preparative (7.8 mm × 30 cm) μBondapak C-18 column (Waters), globin chains from 200 μg to 5.0 mg have been isolated in pure forms. Analytical and preparative procedures using short (50-min duration) and extended (186-min duration) gradient programs have been developed. In addition to the type of column and developer conditions, the following factors are found to be important: (a) preparation of sample, (b) sample loading, and (c) cleaning of the column. Preliminary studies indicate that the yield ranges from 40 to 60% depending on the type of globin sample and the age of the column. This procedure also permits the separation of α, β, and various γ globin chains from fetal and adult samples.
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