E. Katona et al., Enzyme-linked immunosorbent assay for the determination of blood coagulation factor XIII A-subunit in plasma and in cell lysates, J IMMUNOL M, 258(1-2), 2001, pp. 127-135
A new one-step ELISA was developed for the determination of the concentrati
on of blood coagulation factor XIII subunit A (FXIII-A) in plasma and in ce
ll lysates. Monoclonal antibodies directed against different epitopes on FX
III-A were used for the assay. The capture antibody was biotinylated on its
carbohydrate moiety and the detection antibody was labelled with horseradi
sh peroxidase. The antigen-antibody reaction was carried out in the well of
a streptavidin-coated microplate. Complex formation with FXIII subunit B (
FXIII-B) and association to fibrinogen did not influence the accessibility
of the antibodies to FXIII-A. The method could be performed within 2 h and
demonstrated good reproducibility, recovery and sensitivity. Plasma samples
could be assayed after storage at - 20 degreesC for at least 6 months. How
ever, in the case of platelet lysates freezing and rethawing resulted in a
significant loss of FXIII-A. FXIII-A concentrations measured in the plasma
samples of healthy individuals and patients correlated well with the concen
trations of complexed plasma FXIII (A(2)B(2)) and with the results of FXIII
activity measurements. A reference range of 46-82 fg/platelet was establis
hed for platelet FXIII-A. (C) 2001 Elsevier Science B.V. All rights reserve
d.