用于白三烯的灵敏且特异的放射免疫测定法的研发。

The development of sensitive and specific radioimmunoassays for leukotrienes.

作者信息

Rokach J, Hayes E C, Girard Y, Lombardo D L, Maycock A L, Rosenthal A S, Young R N, Zamboni R, Zweerink H J

出版信息

Prostaglandins Leukot Med. 1984 Jan;13(1):21-5. doi: 10.1016/0262-1746(84)90098-2.

Abstract

Radioimmunoassays for leukotriene C4 (LTC4) and for leukotriene B4 (LTB4) have been developed. LTC4 was conjugated with thiolated hemocyanin (Keyhole Limpet) (KLH) using 6-(N-maleimido)hexanoic acid chloride as coupling agent. LTB4 was converted to its hydrazide derivative, via the delta-lactone and the hydrazide was similarly coupled with thiolated KLH using 6-(N-maleimido)hexanoic acid chloride as coupling agent. These conjugates were used to consistently raise high titres of anti-leukotriene antibodies in rabbits. 14,15-[3H]-LTC4 was prepared by total synthesis via two routes. 14,15-[3H]-LTB4 was prepared by total synthesis. The assay for LTC4 recognizes LTC4, LTD4 and LTF4, and to a lesser extent, LTE4 with a detection limit of ca. 0.1 pmoles LTC4 per mL of sample. The assay for LTB4 is highly specific and has a similar detection limit.

摘要

已开发出白三烯C4（LTC4）和白三烯B4（LTB4）的放射免疫分析方法。使用6-（N-马来酰亚胺基）己酰氯作为偶联剂，将LTC4与硫醇化血蓝蛋白（钥孔戚血蓝蛋白）（KLH）偶联。LTB4通过δ-内酯转化为其酰肼衍生物，同样使用6-（N-马来酰亚胺基）己酰氯作为偶联剂，将酰肼与硫醇化KLH偶联。这些偶联物用于在兔体内持续产生高滴度的抗白三烯抗体。14,15-[3H]-LTC4通过两条路线全合成制备。14,15-[3H]-LTB4通过全合成制备。LTC4分析方法可识别LTC4、LTD4和LTF4，在较小程度上还可识别LTE4，检测限约为每毫升样品0.1皮摩尔LTC4。LTB4分析方法具有高度特异性，且检测限相似。