A picogram BA-ELISA quantification assay for rLj-RGD3, a platelet fibrinogen receptor antagonist, in the rat plasma and its application to a pharmacokinetic study.

IF 3.8 2区医学 Q1 Medicine

PLoS Neglected Tropical Diseases Pub Date : 2023-08-17 eCollection Date: 2023-08-01 DOI:10.1371/journal.pntd.0011568

Yuping Wang, Zhien Liu, Guozhu Han, Ping Yu, Xiaobo Yang, Jihong Wang, Li Lv

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引用次数: 0

Abstract

rLj-RGD3, a new member of the RGD (Arginine-Glycine-Aspartate)-motif toxin protein family obtained from Lampetra japonica by means of recombinant DNA techniques, has been demonstrated to be a platelet fibrinogen receptor antagonist and holds potential as a drug candidate for a specific indication. The present article reports an innovative validated highly sensitive and specific biotin-avidin enzyme linked immunosorbent assay (BA-ELISA) to provide a bio-analytical method for pharmacokinetic (PK) studies of rLj-RGD3. The concentration of picogram level rLj-RGD3 in rat plasma was measured using the developed double sandwich BA-ELISA assay, which used two mouse anti-rLj-RGD3 monoclonal antibodies that recognize different epitopes for capture and detection. This method was verified to be highly specific (blank plasma did not interfere with detection), precise (RSD <15%), and accurate (86%-113%). Absolute recovery was in the 94%-119% range. The calibration curve showed good linearity within the 50 to 1600 pg/mL range. The LOQ was as low as 50 pg/mL. The above validated assay was successfully employed to assess PK of rLj-RGD3 in rats. After i.v. and s.c. dosing with 30 μg/kg, the rLj-RGD3 plasma concentration declined bi-exponentially with time. This decay was best fitted to a two-compartment model. In conclusion, the BA-ELISA method described here meets all requirements for PK studies of rLj-RGD3 with an effective pharmacological dose in the μg/kg BW range.

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大鼠血浆中血小板纤维蛋白原受体拮抗剂rLj-RGD3的皮克BA-ELISA定量分析及其在药代动力学研究中的应用。

rLj-RGD3是通过重组DNA技术从日本七孔虫中获得的RGD（精氨酸-甘氨酸-天冬氨酸）基序毒素蛋白家族的新成员，已被证明是血小板纤维蛋白原受体拮抗剂，并具有作为特定适应症候选药物的潜力。本文报道了一种创新的、经验证的高灵敏度和特异性的生物素-亲和素酶联免疫吸附测定法（BA-ELISA），为rLj-RGD3的药代动力学（PK）研究提供了一种生物分析方法。使用开发的双夹心BA-ELISA测定法测量大鼠血浆中皮克水平rLj-RGD3的浓度，该测定法使用两种识别不同表位的小鼠抗rLj-RGB单克隆抗体进行捕获和检测。该方法经验证具有高度特异性（空白血浆不干扰检测）、精密度（RSD

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来源期刊

PLoS Neglected Tropical Diseases Medicine-Infectious Diseases

CiteScore

7.40

自引率

10.50%

发文量

723

审稿时长

2-3 weeks

期刊介绍： PLOS Neglected Tropical Diseases publishes research devoted to the pathology, epidemiology, prevention, treatment and control of the neglected tropical diseases (NTDs), as well as relevant public policy. The NTDs are defined as a group of poverty-promoting chronic infectious diseases, which primarily occur in rural areas and poor urban areas of low-income and middle-income countries. Their impact on child health and development, pregnancy, and worker productivity, as well as their stigmatizing features limit economic stability. All aspects of these diseases are considered, including: Pathogenesis Clinical features Pharmacology and treatment Diagnosis Epidemiology Vector biology Vaccinology and prevention Demographic, ecological and social determinants Public health and policy aspects (including cost-effectiveness analyses).