Development of a sensitive and specific radioreceptor assay for leukotriene B/sub 4/

Journal Article · Fri Jan 01 04:00:00 EST 1988 · Life Sci.; (United States)

DOI:https://doi.org/10.1016/0024-3205(88)90376-1· OSTI ID:7253533

Kohi, F; Agrawal, D K; Cheng, J B; Bewtra, A; Townley, R G; Olesch, J W

To establish a simple and sensitive quantitation of leukotriene B/sub 4/ (LTB/sub 4/), they developed a radioreceptor assay (RRA) using a highly specific (/sup 3/H)leukotriene B/sub 4/((/sup 3/H)LTB/sub 4/) binding to a guinea pig spleen homogenate. The assay detected LTB/sub 4/ levels as low as 0.12 pmol per tube. 50% inhibition of bound (/sup 3/H)LTB/sub 4/ was obtained by 2.5 nM of unlabeled LTB/sub 4/. (/sup 3/H)LTB/sub 4/ competition studies indicated that 20-hydroxy-LTB/sub 4/ was 8 times, 6 trans-LTB/sub 4/ was 640 times and 20-carboxy-LTB/sub 4/ was 1000 times less effective than LTB/sub 4/. The peptide leukotrienes C/sub 4/, D/sub 4/ and E/sub 4/ showed no effect on (/sup 3/H)LTB/sub 4/ binding. Recovery rates averaged 97% after ethanol extraction and evaporation of known amounts of LTB/sub 4/. The intra-assay coefficients of variation for three samples were 2.4%, 7.2% and 8.4%, respectively. This assay was validated by measuring LTB/sub 4/ released from human granulocytes stimulated with calcium ionophore A23187. The LTB/sub 4/ level was maximal at 10 min and decreased rapidly after 15 min. This radioreceptor assay for leukotriene B/sub 4/ is highly sensitive and is comparable to the reported sensitivity by radioimmunoassay. The method is simpler and less expensive than other methods such as high pressure liquid chromatography and is suitable for routine measurement of leukotriene B/sub 4/.

• Research Organization: Creighton Univ. School of Medicine, Omaha, NE (USA)
• OSTI ID: 7253533
• Journal Information: Life Sci.; (United States), Journal Name: Life Sci.; (United States) Vol. 42:22; ISSN LIFSA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ANIMALS
ARACHIDONIC ACID
BIOCHEMICAL REACTION KINETICS
BODY
CARBOXYLIC ACIDS
GUINEA PIGS
HOMOGENATES
ISOTOPE APPLICATIONS
KINETICS
LABELLED COMPOUNDS
MAMMALS
MONOCARBOXYLIC ACIDS
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANS
RADIORECEPTOR ASSAY
REACTION KINETICS
RODENTS
SENSITIVITY ANALYSIS
SPLEEN
TRACER TECHNIQUES
TRITIUM COMPOUNDS
VERTEBRATES