Establishment of 11-dehydro-thromboxane B2 time-resolved immunoassay and application in membranous nephropathy,Analytical Biochemistry

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Establishment of 11-dehydro-thromboxane B2 time-resolved immunoassay and application in membranous nephropathy
Analytical Biochemistry Pub Date : 2023-07-23 , DOI: 10.1016/j.ab.2023.115252
Xuxiang Hui ₁ , Qiuhua Zhang ₂ , Jiayu Li ₁ , Yuan Qin ₁ , Xiumei Zhou ₁ , Xueqin Zhao ₁ , Yan Xu ₃ , Biao Huang ₁

Affiliation

Background

11-Dehydro-thromboxane B2 (11-dehydro-TXB2) is the final stable metabolite of thromboxane A2 (TXA2) and is involved in thrombus formation. Patients with membranous nephropathy (MN) are prone to thromboembolism events.

Methods

Time-resolved fluorescence immunoassay (TRFIA) for 11-dehydro-TXB2 was established by indirect competitive method. The coated 11-dehydro-TXB2-BSA conjugate was used to bind the 11-dehydro-TXB2 antibody competitively to the 11-dehydro-TXB2 antigen in the samples, followed by Eu³⁺-labeled goat anti-mouse IgG antibody, to detect 11-dehydro-TXB2. This study measured 11-dehydro-TXB2 concentrations in serum samples from healthy individuals and patients with MN.

Results

The linear range of TRFIA was 16.38–2000 pg/mL, the sensitivity was 4.70 pg/mL, the average coefficients of variation from intra-assay and inter-assay were 3.50% and 4.95%, respectively, and the recovery was 99.38%. The serum level of 11-dehydro-TXB2 in patients with MN was significantly higher than that in healthy subjects (P < 0.05). The serum 11-dehydro-TXB2 concentration detected by TRFIA was highly consistent with that by ELISA (ρ = 0.900).

Discussion

This study successfully established a new highly sensitive method for the detection of 11-dehydro-TXB2 in serum. 11-Dehydro-TXB2 has great potential in evaluating the risk of thromboembolic events in patients with MN and is expected to be applied to other thromboembolic-related diseases.

中文翻译：

11-脱氢血栓烷B2时间分辨免疫分析方法的建立及其在膜性肾病中的应用

背景

11-脱氢-血栓素 B2 (11-脱氢-TXB2) 是血栓素 A2 (TXA2) 的最终稳定代谢物，参与血栓形成。膜性肾病（MN）患者容易发生血栓栓塞事件。

方法

采用间接竞争法建立了11-脱氢-TXB2的时间分辨荧光免疫分析（TRFIA）。包被的11-脱氢-TXB2-BSA缀合物用于使11-脱氢-TXB2抗体与样品中的11-脱氢-TXB2抗原竞争性结合，然后与Eu ³⁺标记的羊抗小鼠IgG抗体结合，检测11-脱氢-TXB2。本研究测量了健康个体和 MN 患者血清样本中 11-脱氢-TXB2 的浓度。

结果

TRFIA的线性范围为16.38~2000 pg/mL，灵敏度为4.70 pg/mL，批内和批间平均变异系数分别为3.50%和4.95%，回收率为99.38%。MN患者血清11-脱氢-TXB2水平显着高于健康受试者（P < 0.05）。TRFIA检测的血清11-脱氢-TXB2浓度与ELISA检测结果高度一致（ρ=0.900）。