Reinke M, Hoppe U, Röder T, Bestmann H J, Mollenhauer J, Brune K
Department of Pharmacology, University of Erlangen-Nürnberg, F.R.G.
Biochim Biophys Acta. 1991 Feb 5;1081(3):274-8. doi: 10.1016/0005-2760(91)90282-m.
A monoclonal antibody (1A-LDR1) against sulfidopeptide leukotrienes (LT) is described. The mAb shows a nearly identical detection limit of about 0.04 ng for LTC4, LTD4, LTE4 and NacLTE4 in standard fluid phase RIA. Steric modifications, however, diminish the sensitivity, as determined for the examples 5-epi-LTC4, 6-epi-LTC4, 5,6-epi-LTC4 and 11-trans-LTC4. No crossreactivity could be observed for LTB4. Crossreactions with components of the LT peptide chain such as L-cysteine or glutathione, as well as with arachidonic acid, were not detectable. In assessing the accuracy of the LT-RIA, recovery experiments with supernatants of mouse peritoneal macrophages and incubates of gastric mucosa showed a good correlation of r = 0.993 and 0.990, respectively. Results of an inhibition experiment with mouse peritoneal macrophages, incubated with several concentrations of indomethacin and nordihydroguaiaretic acid (NDGA), support the reliability of RIA and ELISA. The new LT-mAB allows an almost complete detection of peptide leukotrienes in one assay.
描述了一种针对硫肽白三烯(LT)的单克隆抗体(1A-LDR1)。在标准液相放射免疫分析中,该单克隆抗体对LTC4、LTD4、LTE4和NacLTE4的检测限几乎相同,约为0.04 ng。然而,空间修饰会降低灵敏度,如对5-表-LTC4、6-表-LTC4、5,6-表-LTC4和11-反式-LTC4的测定所示。未观察到对LTB4的交叉反应。未检测到与LT肽链成分如L-半胱氨酸或谷胱甘肽以及花生四烯酸的交叉反应。在评估LT-放射免疫分析的准确性时,对小鼠腹腔巨噬细胞上清液和胃黏膜孵育物进行的回收率实验分别显示出良好的相关性,r值分别为0.993和0.990。用几种浓度的吲哚美辛和去甲二氢愈创木酸(NDGA)孵育小鼠腹腔巨噬细胞的抑制实验结果支持了放射免疫分析和酶联免疫吸附测定的可靠性。这种新的LT-单克隆抗体能够在一次检测中几乎完全检测出肽白三烯。