Based On The Immunogenicity Of The Hemoglobin Of Blood Products And Safety Study

To investigate the differences of the immunogenic properties at animal study level and antigenic structural or property level between human hemoglobin (hHb), bovine hemoglobin (bHb), porcine hemoglobin (pHb) and their related glutaraldehyde-cross-linked derivatives.Repeated sc administration of an immunizing dose of rabbits' glutaraldehyde-cross-linked derivatives was given to rabbits to stimulate IgG response with an adjuvant. Meanwhile, at the absence of an adjuvant, repeated sc administration of an immunizing dose of hHb,bHb,pHb or their glutaraldehyde-cross-linked derivatives was given to mice to stimulate IgG responses, which was semi-quantitatively measured with an indirect enzyme-linked immunosorbent assay (ELISA). Amino acid sequence comparison between hemoglobin from the three species, western blotting analysis of the antibody-antigen cross-binding profile between glutaradehyde-cross-linked hemoglobin and anti-hemoglobin antibodies raised in mice were used in order to compare the difference in the immunogenic properties, antigenic specificity, physical properties or amino acid sequence of species-specific antigen determinants and/or related submolecular structure in the proteins concerned.In the absence of an adjuvant, hHb exhibited very weak IgG-stimulating effects in mice, while repeated sc administration of bHb or pHb failed to elicit detectable specific IgG response in the animals. and glutaraldehyde polymerized rabbits' hemoglobin failed to elicit detectable specific IgG in the rabbits with an adjuvant too. However, human, bovine, porcine all glutaraldehyde-cross-linked hemoglobin stimulated significant IgG production. The effect of porcine glutaraldehyde polymerized hemoglobin (pPolyHb) was weaker than that of human glutaraldehyde polymerized hemoglobin (hPolyHb) or bovine glutaraldehyde polymerized hemoglobin (bPolyHb). The IgG-stimulating effect of hPolyHb was comparable or slightly higher than that of bPolyHb. The antibody-antigen cross-binding experiences revealed that anti-porcine hemoglobin PcAb may combine bovine hemoglobin,but other mice anti-hemoglobin PcAb hadn't cross-binding response with relevant PolyHb.There were no neo-antigens in glutaraldehyde polymerized hemoglobin. hHb, bHb and pHb are all weakly immunogenic, while polymerization significantly increases the immunogenicity of all their glutaraldehyde-cross-linked derivatives, among which pPolyHb is the weakest. hPolyHb is similar or slightly stronger than bPolyHb in stimulating IgG response in mice. There exist clear species-specific antigen-determinants in hHb, bHb and pHb. N₁₀₈VLVC₁₁₂ is an amino acid sequence of a hHb-specific antigen determinant with C₁₁₂ as the crucial amino acid residue indetermining of its specie specificity o While bHb and pHb possess independent species-specific antigen determinants, they also share some antigen determinants, which do not exist in hHb. However, at a mouse immunological background, bHb and pHb exhibits properties of completely different, immunogenically-active antigen determinant(s).The recognition of differences and homogies in the immunogenic properties at animal study level ,which have important preferences value for researching based porcine glutaraldehyde polymerized hemoglobin HBOCs, and have important function in biological and clinical research based hemoglobin HBOCs.