Payette P J, Cormier M, Dabek B, Yungblut P, Presseault S, Climie S, Sahai J, Cameron W D, Filion L G
Department of Microbiology and Immunology, University of Ottawa, Faculty of Medicine, Ontario, Canada.
Clin Diagn Lab Immunol. 1997 Nov;4(6):671-5. doi: 10.1128/cdli.4.6.671-675.1997.
ALX40-4C is an antiretrovirus agent that has been found to have some inhibitory properties against human immunodeficiency virus (HIV) replication in vitro. The compound was designed as a competitor of the HIV Tat protein for TAR binding. In addition to its anti-HIV properties, it has demonstrated the ability to inhibit in vitro replication of herpes simplex virus types 1 and 2 as well as human cytomegalovirus. Subsequently, in vivo pharmacokinetic evaluation of ALX40-4C necessitated the establishment of a detection system for the measurement of ALX40-4C in subject serum. For this purpose, an indirect-competition enzyme-linked immunosorbent assay with generated rabbit anti-ALX40-4C antiserum was developed. The original assay took 12 h to complete and required many manipulations. Herein, we describe alterations to the system that resulted in the overall reduction in assay time and manipulation. We demonstrate that our alterations do not affect the specificity or sensitivity of the assay compared to that of the original system. ALX40-4C levels in spiked serum samples as well as drug levels from patient samples were used to validate the assay.
ALX40-4C是一种抗逆转录病毒药物，已发现其在体外对人类免疫缺陷病毒（HIV）复制具有一定的抑制特性。该化合物被设计为HIV反式激活因子（Tat）蛋白与反式激活应答元件（TAR）结合的竞争剂。除了具有抗HIV特性外，它还显示出能够在体外抑制1型和2型单纯疱疹病毒以及人巨细胞病毒的复制。随后，对ALX40-4C进行体内药代动力学评估需要建立一个用于测量受试者血清中ALX40-4C的检测系统。为此，开发了一种使用产生的兔抗ALX40-4C抗血清的间接竞争酶联免疫吸附测定法。最初的测定需要12小时完成，且需要许多操作步骤。在此，我们描述了对该系统的改进，这些改进导致测定时间和操作步骤总体减少。我们证明，与原始系统相比，我们的改进不会影响测定的特异性或灵敏度。加标血清样品中的ALX40-4C水平以及患者样品中的药物水平用于验证该测定法。
