Detection of Lewis, P1, and some MNS blood group system antibodies by a solid phase assay

Some solid phase red cell adherence (SPRCA) assays are designed to detect IgG antibodies to red blood cell (RBC) antigens. These assays use anti-IgG-coated red cells as the indicator. It is reported that most antibodies to Lea, Leb, P1, M, and N fail to react by solid phase (SP), presumably because they are IgM antibodies. Those detected are assumed to be IgG. In one year, during routine testing using SPRCA to screen patients for intended RBC transfusion, 28 of 59 such examples were found to react: anti-Lea(9), -Leb(1), -M(14), -N(1), and -P1(3). A study was undertaken to determine if reactivity was due to crosslinking by IgM antibodies of antigen-positive indicator RBCs to antigen-positive reagent RBC monolayers, or due to detection of IgG antibodies. Antibodies were tested according to standard SP protocols, except where IgG-neutralized indicator RBCs were substituted for anti-IgG-active indicator cells. The 59 samples were retested with antigen-positive and antigen-negative indicator RBCs. Only 5 of 59 reacted optimally when antigen-positive indicator cells were used: anti-Lea(2), -Leb(1), -M(1), and -N(1). The reactions of all antibodies were abolished when the anti-IgG component of the indicator was neutralized by soluble IgG. These findings show that detection of most Lewis, P1, M, and N antibodies by SPRCA is dependent on the presence of an IgG antibody in the serum.