Background: Chemical biotinylation of red cell membranes may be useful for several clinical applications, including red cell survival studies.
Study design and methods: To examine the possible effects of biotinylation on red cell antigens, standard hemagglutination assays were performed on matched sets of control and biotinylated red cells. The red cells were biotinylated at a final concentration of 2.0 pg of sulfo-N-hydroxysuccinimide-biotin per cell, and antigen-negative cells were directly compared to antigen-positive cells when possible. The hemagglutination assays were graded in a blinded fashion. Forty-one red cell antigens from 21 of the 23 established blood group systems were tested.
Results: Hemagglutination based upon antibody binding to A, A1, M, N, S, s, P1, D, C, E, c, e, C(w), Lu(b), K, k, Kp(b), Le(a), Le(b), Fy(a), Fy(b), Jk(a), Jk(b), Di(a), Wr(a), Wr(b), Yt(a), Xg(a), Sc1, Do(b), Co(a), Ch, H, Ge2, Cr(a), Kn(a), I, and P was not affected by biotinylation. Unexpectedly, the hemagglutination of Di(b+) and LW(a+) red cells was blocked after biotinylation. Conversely, MH04 monoclonal anti-A agglutinated red cells expressing B only after biotinylation. BIRMA-1 monoclonal anti-A and polyclonal anti-A from sera did not agglutinate the biotinylated B red cells.
Conclusion: Biotinylation of human red cells specifically modified their antigenicity, as measured by standard hemagglutination assays.