通过高效液相色谱法和放射免疫分析法联合使用检测白三烯的生物合成与代谢。

Leukotriene biosynthesis and metabolism detected by the combined use of HPLC and radioimmunoassay.

作者信息

Beaubien B C, Tippins J R, Morris H R

出版信息

Biochem Biophys Res Commun. 1984 Nov 30;125(1):97-104. doi: 10.1016/s0006-291x(84)80339-3.

Abstract

A radioimmunoassay for leukotriene D4 (LTD4) has been developed which exhibits sufficiently high sensitivity to be useful in conjunction with RP-HPLC in the detection of LTC4, LTD4 and LTE4 in physiological samples. The detection limit of the assay was approximately 240 amoles, using antiserum TG1 at a dilution of 6 X 10(3), with 50% displacement at 70 fmoles. Antiserum NW1, also at a dilution of 6 X 10(3), displayed a detection limit of 9 fmoles with 50% displacement at 100 fmoles. The two antisera have similiar crossreactivities, both manifesting useful affinities for LTE4 and LTC4, and low or negligible affinities for other arachidonic acid metabolites, or their derivatives. The radioimmunoassay was used to detect 1) LTC4, LTD4 and LTE4 released from perfused rat lung in response to platelet-activating factor (PAF) stimulation, 2) conversion of exogenous LTD4 to LTE4 in human blood, and 3) endogenous leukotrienes in human blood samples.

摘要

已开发出一种白三烯D4（LTD4）放射免疫测定法，该方法具有足够高的灵敏度，可与反相高效液相色谱法（RP-HPLC）结合用于检测生理样品中的LTC4、LTD4和LTE4。使用稀释度为6×10³的抗血清TG1时，该测定法的检测限约为240阿托摩尔，在70飞摩尔时50%被置换。同样稀释度为6×10³的抗血清NW1，检测限为9飞摩尔，在100飞摩尔时50%被置换。这两种抗血清具有相似的交叉反应性，对LTE4和LTC4均表现出有用的亲和力，对其他花生四烯酸代谢产物或其衍生物的亲和力较低或可忽略不计。该放射免疫测定法用于检测：1）灌注大鼠肺在血小板活化因子（PAF）刺激下释放的LTC4、LTD4和LTE4；2）人血中外源性LTD4向LTE4的转化；3）人血样品中的内源性白三烯。