The development of a sensitive and specific radioreceptor assay for leukotriene B4

F. Kohi, Devendra K. Agrawal, J. B. Cheng, Againdra K. Bewtra, R. G. Townley, J. W. Olesch

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

To establish a simple and sensitive quantitation of leukotriene B4 (LTB4), we developed a radioreceptor assay (RRA) using a highly specific [3H] leukotriene B4([3H]LTB4) binding to a guinea pig spleen homogenate. The assay detected LTB4 levels as low as 0.12 pmol per tube. Fifty percent inhibition of bound [3H]LTB4 was obtained by 2.5 nM of unlabeled LTB4. [3H]LTB4 competition studies indicated that 20-hydroxy-LTB4 was 8 times, 6-trans-LTB4 was 640 times and 20-carboxy-LTB4 was 1000 times less effective than LTB. The peptide leukotrienes C4, D4 and E4 showed no effect on [3H]LTB4 binding. Recovery rates averaged 97% after ethanol extraction and evaporation of known amounts of LTB4. The intra-assay coefficients of variation for three samples were 2.4%, 7.2% and 8.4%, respectively. This assay was validated by measuring LTB4 released from human granulocytes stimulated with calcium ionophore A23187. The LTB4 level was maximal at 10 min (156.8 ± 36.2 pmol/3×106 cells) and decreased rapidly after 15 min. This radioreceptor assay for leukotriene B4 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 B4.

Original languageEnglish
Pages (from-to)2241-2248
Number of pages8
JournalLife Sciences
Volume42
Issue number22
DOIs
StatePublished - 1988

Fingerprint

Radioligand Assay
Leukotriene B4
Assays
High pressure liquid chromatography
Leukotriene E4
Leukotriene D4
Leukotriene C4
Calcium Ionophores
Calcimycin
Granulocytes
Radioimmunoassay

All Science Journal Classification (ASJC) codes

  • Pharmacology

Cite this

The development of a sensitive and specific radioreceptor assay for leukotriene B4 . / Kohi, F.; Agrawal, Devendra K.; Cheng, J. B.; Bewtra, Againdra K.; Townley, R. G.; Olesch, J. W.

In: Life Sciences, Vol. 42, No. 22, 1988, p. 2241-2248.

Research output: Contribution to journalArticle

Kohi, F. ; Agrawal, Devendra K. ; Cheng, J. B. ; Bewtra, Againdra K. ; Townley, R. G. ; Olesch, J. W. / The development of a sensitive and specific radioreceptor assay for leukotriene B4 . In: Life Sciences. 1988 ; Vol. 42, No. 22. pp. 2241-2248.
@article{4190d860684b40fa8db5f09ea6713e0a,
title = "The development of a sensitive and specific radioreceptor assay for leukotriene B4",
abstract = "To establish a simple and sensitive quantitation of leukotriene B4 (LTB4), we developed a radioreceptor assay (RRA) using a highly specific [3H] leukotriene B4([3H]LTB4) binding to a guinea pig spleen homogenate. The assay detected LTB4 levels as low as 0.12 pmol per tube. Fifty percent inhibition of bound [3H]LTB4 was obtained by 2.5 nM of unlabeled LTB4. [3H]LTB4 competition studies indicated that 20-hydroxy-LTB4 was 8 times, 6-trans-LTB4 was 640 times and 20-carboxy-LTB4 was 1000 times less effective than LTB. The peptide leukotrienes C4, D4 and E4 showed no effect on [3H]LTB4 binding. Recovery rates averaged 97{\%} after ethanol extraction and evaporation of known amounts of LTB4. The intra-assay coefficients of variation for three samples were 2.4{\%}, 7.2{\%} and 8.4{\%}, respectively. This assay was validated by measuring LTB4 released from human granulocytes stimulated with calcium ionophore A23187. The LTB4 level was maximal at 10 min (156.8 ± 36.2 pmol/3×106 cells) and decreased rapidly after 15 min. This radioreceptor assay for leukotriene B4 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 B4.",
author = "F. Kohi and Agrawal, {Devendra K.} and Cheng, {J. B.} and Bewtra, {Againdra K.} and Townley, {R. G.} and Olesch, {J. W.}",
year = "1988",
doi = "10.1016/0024-3205(88)90376-1",
language = "English",
volume = "42",
pages = "2241--2248",
journal = "Life Sciences",
issn = "0024-3205",
publisher = "Elsevier Inc.",
number = "22",

}

TY - JOUR

T1 - The development of a sensitive and specific radioreceptor assay for leukotriene B4

AU - Kohi, F.

AU - Agrawal, Devendra K.

AU - Cheng, J. B.

AU - Bewtra, Againdra K.

AU - Townley, R. G.

AU - Olesch, J. W.

PY - 1988

Y1 - 1988

N2 - To establish a simple and sensitive quantitation of leukotriene B4 (LTB4), we developed a radioreceptor assay (RRA) using a highly specific [3H] leukotriene B4([3H]LTB4) binding to a guinea pig spleen homogenate. The assay detected LTB4 levels as low as 0.12 pmol per tube. Fifty percent inhibition of bound [3H]LTB4 was obtained by 2.5 nM of unlabeled LTB4. [3H]LTB4 competition studies indicated that 20-hydroxy-LTB4 was 8 times, 6-trans-LTB4 was 640 times and 20-carboxy-LTB4 was 1000 times less effective than LTB. The peptide leukotrienes C4, D4 and E4 showed no effect on [3H]LTB4 binding. Recovery rates averaged 97% after ethanol extraction and evaporation of known amounts of LTB4. The intra-assay coefficients of variation for three samples were 2.4%, 7.2% and 8.4%, respectively. This assay was validated by measuring LTB4 released from human granulocytes stimulated with calcium ionophore A23187. The LTB4 level was maximal at 10 min (156.8 ± 36.2 pmol/3×106 cells) and decreased rapidly after 15 min. This radioreceptor assay for leukotriene B4 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 B4.

AB - To establish a simple and sensitive quantitation of leukotriene B4 (LTB4), we developed a radioreceptor assay (RRA) using a highly specific [3H] leukotriene B4([3H]LTB4) binding to a guinea pig spleen homogenate. The assay detected LTB4 levels as low as 0.12 pmol per tube. Fifty percent inhibition of bound [3H]LTB4 was obtained by 2.5 nM of unlabeled LTB4. [3H]LTB4 competition studies indicated that 20-hydroxy-LTB4 was 8 times, 6-trans-LTB4 was 640 times and 20-carboxy-LTB4 was 1000 times less effective than LTB. The peptide leukotrienes C4, D4 and E4 showed no effect on [3H]LTB4 binding. Recovery rates averaged 97% after ethanol extraction and evaporation of known amounts of LTB4. The intra-assay coefficients of variation for three samples were 2.4%, 7.2% and 8.4%, respectively. This assay was validated by measuring LTB4 released from human granulocytes stimulated with calcium ionophore A23187. The LTB4 level was maximal at 10 min (156.8 ± 36.2 pmol/3×106 cells) and decreased rapidly after 15 min. This radioreceptor assay for leukotriene B4 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 B4.

UR - http://www.scopus.com/inward/record.url?scp=0023942355&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0023942355&partnerID=8YFLogxK

U2 - 10.1016/0024-3205(88)90376-1

DO - 10.1016/0024-3205(88)90376-1

M3 - Article

C2 - 2836680

AN - SCOPUS:0023942355

VL - 42

SP - 2241

EP - 2248

JO - Life Sciences

JF - Life Sciences

SN - 0024-3205

IS - 22

ER -