Background: Hemagglutination for detection and semiquantification of ABO antibodies is associated with large center-to-center variations and poor reproducibility. Because acceptance for transplantation and diagnosis of rejection in ABO-incompatible transplantation rely on the levels and specificity of ABO antibodies, reproducible tests that allow their detection and specificity determination are required.
Study design and methods: The level of chain type-specific anti-A and anti-B were analyzed in the sera of 44 healthy individuals of known ABO blood group using an enzyme-linked immunosorbent assay (ELISA) with polyacrylamide (PAA) conjugates of blood group A and B trisaccharides or Type 2 chain A and B tetrasaccharides. Selected sera were further analyzed by hemagglutination and in an ELISA with Types 1 to 4 chain A or B neoglycolipids (NGL) as antigens.
Results: Immunoglobulin (Ig)G anti-A and anti-B levels were higher (p ≤ 0.05) in blood group O than in B and A individuals. More IgM anti-A and anti-B cross-reactivity was detected in AB serum on PAA-conjugated A and B trisaccharides than on the tetrasaccharides. One of 11 blood group B and two of 12 A individuals had IgG antibodies binding the tetrasaccharide despite lack of, or very low reactivity with, the trisaccharides. IgG antibodies preferring the A and B Type 2 tetrasaccharides were of the IgG2 subclass. The NGL ELISA further supported the presence of chain type-specific anti-A and -B antibodies among nonsensitized, healthy individuals.
Conclusion: An ELISA with structurally defined ABH antigens will allow the antibody class and fine specificity of ABO antibodies to be determined, which may improve risk assessment in ABO-incompatible transplantation.
© 2010 American Association of Blood Banks.