PRODUCTION AND CHARACTERIZATION OF MONOCLONAL-ANTIBODIES THAT SPECIFICALLY BIND TO PHOSPHATIDYLCHOLINE
PRODUCTION AND CHARACTERIZATION OF MONOCLONAL-ANTIBODIES THAT SPECIFICALLY BIND TO PHOSPHATIDYLCHOLINE
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DOI：
10.1016/0005-2760(90)90098-i
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发表时间：
1990-08-28
期刊：
影响因子：
--
通讯作者：
INOUE, K
中科院分区：
文献类型：
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作者：
NAM, KS;IGARASHI, K;INOUE, K
建立了一系列与磷脂酰胆碱（PC）反应的单克隆抗体（mAb）。所有mAb均对PC具有高度特异性，且未观察到与其他磷脂的交叉反应。描述了用两种典型的单克隆抗体JE-1和JE-8所获得的结果。使用具有改性极性头基的合成PC类似物的分析表明，胆碱部分的季氮上的甲基对于结合是重要的。每种单克隆抗体显示出PC分子的不同的酰基链特异性，并且JE-1显示出与具有饱和脂肪酸的PC相当大的反应性，而JE-8不能与PC反应。两种mAb均与具有不饱和脂肪酸的PC结合，但显示出不同的反应性特征。两种mAb仅与水溶性半抗原如磷酸胆碱和L-α-半抗原弱反应。甘油磷酸胆碱，表明PC分子的疏水部分是重要的最大亲和力。通过分析单克隆抗体对磷脂酶A2和磷脂酶C活性的影响，进一步研究了单克隆抗体与PC分子疏水部分的相互作用。JE-1抑制两种酶的活性，而JE-8仅抑制磷脂酶C的活性，表明JE-1比JE-8更彻底地与PC分子的疏水区域相互作用。
A series of monoclonal antibodies (mAbs) that react with phosphatidylcholine (PC) were established. All mAbs were highly specific to PC and no cross-reaction with other phospholipids was observed. The results obtained with two typical monoclonal antibodies, JE-1 and JE-8, were described. The analysis using synthetic PC analogs with modified polar head groups showed that the methyl groups on the quaternary nitrogen of the choline moiety were important for the binding. Each mAbs showed distinct acyl chain specificities of the PC molecules, and JE-1 showed considerable reactivity with PC with saturated fatty acids, whereas JE-8 could not react with the PC. Both mAbs bound to PC with unsaturated fatty acids, but showed distinct reactivity profiles. Both mAbs reacted only weakly with water-soluble haptens such as phosphorylcholine and L-.alpha.-glycerophosphocholine, suggesting that the hydrophobic moiety of the PC molecule is important for the maximum affintity. The interaction between the mAbs and the hydrophobic moieties of PC molecules was further studied by analyzing the effect of the mAbs on the activities of phospholipase A2 and phospholipase C. JE-1 inhibited both enzyme activities, while JE-8 inhibited only the phospholipase C activity, indicating that JE-1 interacts more thoroughly with the hydrophobic region of the PC molecule than JE-8 does.