Hemoglobin Structure Changes and Their Impact on Function

Abstract

Detailed studies on the abnormal hemoglobin from antenatal pregnant women have been undertaken to determine the distribution of abnormal hemoglobin and their relation to anemic condition during pregnancy. Blood samples (1050) were collected in EDTA, from Antenatal Clinic, Department of Obstetric and Gynecology, Jinnah Postgraduate Medical Centre, Karachi. Whole blood hemoglobin concentration was determined according to Drabkins. This study shows that about 70% in this group were anemic. The screening of abnormal hemoglobin was carried out electrophoretically on Cellulose acetate membrane in (TEB) Tris.EDTA.Borate buffer. It shows that 20 samples are abnormal of which 7 seems to be new variant. Separation of globin chains was achieved on CM. Cellulose in 8-Urea and also by Reversed Phase H.P.L.C. by using column of Water-Rpl8 μ-Bonda pak with 0.1% TFA and increasing acetonitrile gradient from (0-60%). Purity of chains was checked by Triton-X-PAG-electrophoresis. Enzymatic digestion of globin chain was done with Trypsin-TPCK. Tryptic peptides were chromatographed by gel filtration on Sephandex G-25 Coarse, and rechromatographed on HPLC. Purity of peptides was checked by thin layer of electrophoresis. Studies on amino acid sequence by DABTIC method are in progress.