Radioimmunoassay for 11-dehydro-TXB2: a method for monitoring thromboxane production in vivo. 1986

M Kumlin, and E Granström

A radioimmunoassay was developed for 11-dehydro-TXB2, a prominent metabolite of TXB2 in blood and urine of several species. In order to reliably assay 11-dehydro-TXB2, its chemical stability as well as its chromatographic properties were first examined. Since dehydrogenation at C-11 converts the thromboxane ring into the delta-lactone form of a dicarboxylic acid, which can also occur in an open form, the analysis of 11-dehydro-TXB2 may be somewhat complicated. In some chromatographic systems, the compound thus migrated with pronounced tailing, and during extraction using the common Sep-Pak procedure the two forms were partially separated. The lactone as well as the open form could be conclusively identified using mass spectrometry. The equilibrium between the two forms of 11-dehydro-TXB2 was studied in buffers of different pH and in plasma. Higher pH favoured hydrolysis into the acyclic structure. The lactonization and hydrolysis processes were also shown to be time and temperature dependent. Two different antiplasms, raised in rabbits against conjugates of 11-dehydro-TXB2 with bovine serum albumin, displayed somewhat different properties in their recognition of the two forms of 11-dehydro-TXB2. A radioimmunoassay employing these antibodies was developed. The labeled antigen was prepared by incubation of 3H-TXB2 with rabbit lung supernatant. The limit of detection was 1.5 pg. For validation of the assay, analysis of blood and urinary samples, obtained after injection of TXB2 to a human volunteer, was done. The values obtained were compatible with previous isotope studies. Results from an inhibition experiment with rabbit lung incubated in the presence or absence of indomethacin further supported the identity of the assayed substance.

UI MeSH Term Description Entries
D008991 Monitoring, Physiologic The continuous measurement of physiological processes, blood pressure, heart rate, renal output, reflexes, respiration, etc., in a patient or experimental animal; includes pharmacologic monitoring, the measurement of administered drugs or their metabolites in the blood, tissues, or urine. Patient Monitoring,Monitoring, Physiological,Physiologic Monitoring,Monitoring, Patient,Physiological Monitoring
D011786 Quality Control A system for verifying and maintaining a desired level of quality in a product or process by careful planning, use of proper equipment, continued inspection, and corrective action as required. (Random House Unabridged Dictionary, 2d ed) Control, Quality,Controls, Quality,Quality Controls
D011817 Rabbits A burrowing plant-eating mammal with hind limbs that are longer than its fore limbs. It belongs to the family Leporidae of the order Lagomorpha, and in contrast to hares, possesses 22 instead of 24 pairs of chromosomes. Belgian Hare,New Zealand Rabbit,New Zealand Rabbits,New Zealand White Rabbit,Rabbit,Rabbit, Domestic,Chinchilla Rabbits,NZW Rabbits,New Zealand White Rabbits,Oryctolagus cuniculus,Chinchilla Rabbit,Domestic Rabbit,Domestic Rabbits,Hare, Belgian,NZW Rabbit,Rabbit, Chinchilla,Rabbit, NZW,Rabbit, New Zealand,Rabbits, Chinchilla,Rabbits, Domestic,Rabbits, NZW,Rabbits, New Zealand,Zealand Rabbit, New,Zealand Rabbits, New,cuniculus, Oryctolagus
D011863 Radioimmunoassay Classic quantitative assay for detection of antigen-antibody reactions using a radioactively labeled substance (radioligand) either directly or indirectly to measure the binding of the unlabeled substance to a specific antibody or other receptor system. Non-immunogenic substances (e.g., haptens) can be measured if coupled to larger carrier proteins (e.g., bovine gamma-globulin or human serum albumin) capable of inducing antibody formation. Radioimmunoassays
D002021 Buffers A chemical system that functions to control the levels of specific ions in solution. When the level of hydrogen ion in solution is controlled the system is called a pH buffer. Buffer
D002845 Chromatography Techniques used to separate mixtures of substances based on differences in the relative affinities of the substances for mobile and stationary phases. A mobile phase (fluid or gas) passes through a column containing a stationary phase of porous solid or liquid coated on a solid support. Usage is both analytical for small amounts and preparative for bulk amounts. Chromatographies
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D006863 Hydrogen-Ion Concentration The normality of a solution with respect to HYDROGEN ions; H+. It is related to acidity measurements in most cases by pH pH,Concentration, Hydrogen-Ion,Concentrations, Hydrogen-Ion,Hydrogen Ion Concentration,Hydrogen-Ion Concentrations
D006868 Hydrolysis The process of cleaving a chemical compound by the addition of a molecule of water.
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia

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