J. Geen et al., Detection of anti-Yt(a) antibodies using a sensitive and specific enzyme-linked immunosorbent assay, BR J BIOMED, 56(2), 1999, pp. 128-133
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.