RECOMBINANT HEMOGLOBIN-A PRODUCED IN TRANSGENIC SWINE - STRUCTURAL EQUIVALENCE WITH HUMAN HEMOGLOBIN-A

Recombinant human hemoglobin A produced by coexpressing human alpha an d beta globin genes in swine, and purified from the lysate of transgen ic swine has been subjected to detailed protein chemical analysis. The se structural studies involving laser desorption mass spectrometry, se paration of globin chains by RPHPLC, amino terminal sequence analysis of the isolated globin chains, the tryptic peptide mapping of the puri fied globin chains and the amino acid composition analysis of the puri fied tryptic peptides of globin chains have established the primary st ructural equivalence of the globin chains of the transgenic swine deri ved hemoglobin A with that of human hemoglobin A. These results demons trate that the transgenic swine system correctly translates the human alpha and beta globin m-RNA; carries out the correct cotranslational p rocessing of globin chains, and does not introduce any unwanted post t ranslational modifications into the mature chains. Thus, the transgeni c swine expression system is an excellent approach for the production of HbA for developing an effective hemoglobin based oxygen carrier.