Journal
JOURNAL OF IMMUNOLOGICAL METHODS
Volume 258, Issue 1-2, Pages 127-135Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/S0022-1759(01)00479-3
Keywords
factor XIII of blood coagulation; factor XIII subunit A; ELISA; platelet; monoclonal antibodies
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A new one-step ELISA was developed for the determination of the concentration of blood coagulation factor XIII subunit A (FXIII-A) in plasma and in cell lysates. Monoclonal antibodies directed against different epitopes on FXIII-A were used for the assay. The capture antibody was biotinylated on its carbohydrate moiety and the detection antibody was labelled with horseradish 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. However, 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 concentrations 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 established for platelet FXIII-A. (C) 2001 Elsevier Science B.V. All rights reserved.
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