Detection of anti-Yt(a) antibodies using a sensitive and specific enzyme-linked immunosorbent assay

A specific, sensitive and semi-quantitative enzyme-linked immunosorbent ass ay (ELISA) is described to detect anti-Yt(a) antibodies in human serum. Rec ombinant;acetylcholinesterase (AChE E.C.3.1.1.7) was employed as the coatin g antigen in the microtitre plate and horseradish peroxidase (HRP)-conjugat ed specific antibody (IgG) was used as the secondary antibody. The method d eveloped showed excellent sensitivity, detecting a titre >1 in 600 000 (3.5 ng/mL mouse IgG protein) for mouse monoclonal (mMAb) anti-AChE antibody. N o cross-reaction was seen with other common blood group antibodies, confirm ing the specificity of the method. The recombinant antigen's AChE phenotype was confirmed as Yt(a), as no reaction errs detected with anti-Yt(b)-posit ive sera. The ELISA method correlated closely with the established serologi cal grading system used routinely in blood transfusion laboratories.