AJP - Endocrinology and Metabolism, Vol 262, Issue 5 E658-E662, Copyright © 1992 by American Physiological Society
Monitoring thromboxane in body fluids: a specific ELISA for 11-dehydrothromboxane B2 using a monoclonal antibody
M. Reinke
Department of Medicine IV, University of Erlangen-Nurnberg, Federal Republic of Germany.
11-Dehydrothromboxane B2 (11-DHTxB2) concentrations are believed to reflect
levels of the in vivo synthesis and release of thromboxane A2. In a
specific enzyme-linked immunosorbent assay (ELISA) a monoclonal antibody
(MAB) against 11-DHTxB2 (MAB-1E-DHTBR1) recognizes the acyclic form of
11-DHTxB2, as found in the basic range of pH, with a detection limit of 4.6
pg/sample and a binding affinity of 6.1 x 10(9) l/mol. Negligible
cross-reactivity was found for thromboxane B2 (0.05%),
2,3-dinor-thromboxane B2 (0.06%), and prostaglandin D2 (0.08%). Validity of
the assay was confirmed by a good correlation between radioimmunoassay and
ELISA (r = 0.972). Recovery experiments showed an accuracy of r = 0.982.
Measurements of 11-DHTxB2 in human serum and urine samples demonstrated the
practical applicability of the MAB in ELISA. With the use of different
clotting times, the serum level of 11-DHTxB2 ranged from 0.8-1.3 ng/ml (30
min) to 24.1-47.9 ng/ml (4 h). After administration of aspirin the
11-DHTxB2 level of human urine declined from 3.9-5.4 ng/ml to 0.4-1.6 ng/ml
after 6 h.
