Wang Juncheng, Liu Meng, Cai Zecheng, Zahid Rukhshan, Zhang Wenjie, Ma Dan, Li Die, Liang Yan, Zha Lei, Zhou Yun, Wang Lina, Yang Gang, Zheng Shuai, Xu Yuekang
Anhui Provincial Key Laboratory for Conservation and Exploitation of Biological Resources, College of Life Sciences, Anhui Normal University, Wuhu, Anhui 241000, China.
The First Affiliated Hospital of Wannan Medical College, Wuhu, Anhui 241001, China.
Int Immunopharmacol. 2024 Dec 25;143(Pt 3):113473. doi: 10.1016/j.intimp.2024.113473. Epub 2024 Nov 15.
Sepsis is a fatal consequence of compromised host immunity due to widespread infection. Its pathogenesis has recently been found to be associated with tetranectin (TN), a monocyte-produced plasma protein with a critical disease-associated epitope, P5-5. To develop a rapid and simple method for early monitoring of the disease in clinical settings, a purified monoclonal antibody (12F1 mAb) with high affinity for the human TN pathogenic epitope P5-5 was produced in this study. The linear range of the indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) based on the mAb to detect TN-P5-5 was 4.8-312 ng/mL, and the half-maximal inhibitory concentration (IC) was 26.99 ng/mL, with a limit of detection of 2.4 ng/mL. Furthermore, the average recovery of intra- and inter-assay were 103.253 ± 2.803 % and 107.778 ± 7.490 %, respectively. Importantly, the competitive ELISA method established using 12F1 revealed signals corresponding to disease severity in patients with sepsis. Furthermore, the specific in vivo recognition of a pathogenic epitope by mAbs can be extended to therapeutic applications. Collectively, the development of an epitope-specific mAb against disease-associated proteins could be utilized accurately and quantitatively for diagnosing and monitoring diseases in clinical blood samples.
脓毒症是广泛感染导致宿主免疫受损的致命后果。最近发现其发病机制与纤连蛋白四聚体（TN）有关，TN是一种由单核细胞产生的血浆蛋白，带有一个与疾病密切相关的表位P5-5。为了开发一种在临床环境中早期监测该疾病的快速简便方法，本研究制备了一种对人TN致病表位P5-5具有高亲和力的纯化单克隆抗体（12F1 mAb）。基于该单克隆抗体检测TN-P5-5的间接竞争酶联免疫吸附测定（ic-ELISA）的线性范围为4.8-312 ng/mL，半数抑制浓度（IC）为26.99 ng/mL，检测限为2.4 ng/mL。此外，批内和批间的平均回收率分别为103.253±2.803%和107.778±7.490%。重要的是，使用12F1建立的竞争ELISA方法揭示了脓毒症患者中与疾病严重程度相对应的信号。此外，单克隆抗体对致病表位的特异性体内识别可扩展到治疗应用。总体而言，针对疾病相关蛋白的表位特异性单克隆抗体的开发可准确、定量地用于临床血样中疾病的诊断和监测。
