The zeta (0 and various gamma (y) chains of Hb Portland I have been separated and isolated from blood samples obtained from neonates with hydrops fetalis due to homozygous a-thalassemia. By using developers containing acetonitrile, methanol, and potassium phosphate and either an analytical (3.9 mm X 30 cm) or a preparative (7.8 mm X 30 cm) PBondapak C-18 column (Waters), globin chains from 200 pg 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 LY, 8. and various y globin chains from fetal and adult samples. Hemoglobin Portland I (Hb Portland)3 was discovered by Capp et al. ( 1) in 1967 in a female Chinese infant with multiple congenital anomalies and a complex autosomal mosaicism. Since then Hb Portland has been reported in infants with hydrops fetalis due to homozygous cr-thalassemia (2, 3), in trace amounts in some normal human cord blood (4), and in hemin-induced K562 cell line cultures (5) along with other embryonic hemoglobins.
Hemoglobin Portland is composed of two alpha-like chains designated zeta (S) chains, and two gamma (y) chains (1,6). Two other human embryonic hemoglobins, Gower 1, first thought to be c4 but later identified as & (7) ' Presented as a poster paper at the International Symposium on HPLC of Proteins and Peptides November