Hayes E C, Lombardo D L, Girard Y, Maycock A L, Rokach J, Rosenthal A S, Young R N, Egan R W, Zweerink H J
J Immunol. 1983 Jul;131(1):429-33.
Rabbits were immunized with leukotriene C4 (LTC4) coupled to thiolated keyhole limpet hemocyanin (KLH) by using 6-N-maleimidohexanoic acid as a spacer molecule. Immune serum was obtained with 7.9 nmol of LTC4-specific immunoglobulin per milliliter and a mean association constant of 2.1 X 10(9) M-1. A radioimmunoassay was developed that detected 0.1 pmol of LTC4 per 1-ml sample. LTD4 and LTE4, three isomers of LTC4, the sulfones of LTC4, LTD4, and LTE4, and one isomer of LTD4 reacted to varying degrees in the assay. A number of other structurally related compounds, such as LTB4 and 5-HETE, did not react. Conditions were established to determine LTC4 levels in human plasma without loss of LTC4 during sample preparation and without the need for extraction procedures before the measurement of LTC4.
通过使用6-N-马来酰亚胺己酸作为间隔分子,将白三烯C4(LTC4)与硫醇化的钥孔血蓝蛋白(KLH)偶联,对兔子进行免疫。获得的免疫血清中每毫升含有7.9 nmol的LTC4特异性免疫球蛋白,平均缔合常数为2.1×10⁹ M⁻¹。开发了一种放射免疫测定法,该方法可检测每1毫升样品中0.1 pmol的LTC4。LTD4和LTE4,LTC4的三种异构体,LTC4、LTD4和LTE4的砜,以及LTD4的一种异构体在该测定中表现出不同程度的反应。许多其他结构相关的化合物,如LTB4和5-HETE,没有反应。建立了在样品制备过程中不损失LTC4且在测量LTC4之前无需提取程序的条件,以测定人血浆中的LTC4水平。