The Major Antigen Of Human, Bovine, Porcine Hemoglobin, And The Corresponding Glutaraldehyde Polymer Epitope Comparative Study

In this article, human, bovine and porcine blood were used as the raw material, ultra-pure human, bovine and porcine hemoglobin (hHb, bHb and pHb) were obtained through multiple purification steps; with glutaraldehyde as the cross-linker, polymerized human, bovine and porcine hemoglobin (hPolyHb, bPolyHb and pPolyHb) were then produced. The immunogenicity of human, bovine, porcine hemoglobin and glutaraldehyde polymerized human, bovine, porcine hemoglobin were investigated through detection of the serum specific IgG responses using indirect ELISA. The methodology of Western Blotting and Ag-Chip was used in this study, with which cross-species antigen/antibody binding reactions among human, bovine, porcine hemoglobin and the related glutaraldehyde polymerized hemoglobin were carried out using polyclonal antibodies against these proteins.The result of indirect ELISA indicated that in the absence of adjuvant, hHb exhibited very weak IgG-stimulating effects in mice, while repeated intraperitoneal administration of bHb or pHb failed to elicit detectable specific IgG response in the animals. However, all glutaraldehyde cross-linked hemoglobin stimulated significant IgG production. The effect of pPolyHb and bPolyHb was weaker than that of hPolyHb. Western Blotting and Ag-Chip results demonstrated that human, bovine and porcine hemoglobin contain distinct antigen determinant, but also share some antigen determinants. However, at mouse or rabbbit immunological background, hHb, bHb and pHb exhibits properties of completely different antigen determinant(s).