Variations in Dominant Antigen Determinants of Glutaraldehyde Polymerized Human, Bovine and Porcine Hemoglobin

Abstract

In this study, immunogenicity of human hemoglobin (hHb), bovine hemoglobin (bHb), porcine hemoglobin (pHb) and their glutaradehyde polymerized derivatives (hPolyHb, bPolyHb and bPolyHb, respectively) were compared. The nature of the dominant antigen determinants of the chemically polymerized proteins was studied. Glutaraldehyde chemical reaction enhanced the immunogenicity of the hemoglobin derivatives. In mice, the extent of the enhancement was largely comparable among hPolyHb, bPolyHb and pPolyHb. Using the methods of semi-quantitative western blotting and quantitative protein array, it was found that most of the polycloncal antibodies raised in rodents against glutaraldehyde polymerized hemoglobin derivatives of human, bovine or porcine species only weakly or did not cross-react with the hemoglobin derivatives of the other two species, indicating that hPolyHb, bPolyHb and bPolyHb vary significantly in their dominant antigen determinants, despite very high degree of identity in their primary amino acid sequences and high similarity in their three dimensional structures.