Development and characterization of three novel mouse monoclonal antibodies targeting spike protein S1 subunit of Middle East Respiratory Syndrome Corona Virus.

Q3 Medicine

Human Antibodies Pub Date : 2024-05-02 DOI:10.3233/HAB-240016

A. Abbas, S. El-Kafrawy, Ashraf A Tabll, A. Hashem, Tagreed L Al Subhi, M. Alsaadi, E. Azhar

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引用次数: 0

Abstract

BACKGROUND Middle East Respiratory Syndrome Coronavirus is a highly pathogenic virus that poses a significant threat to public health. OBJECTIVE The purpose of this study is to develop and characterize novel mouse monoclonal antibodies targeting the spike protein S1 subunit of the Middle East Respiratory Syndrome Corona Virus (MERS-CoV). METHODS In this study, three mouse monoclonal antibodies (mAbs) against MERS-CoV were generated and characterized using hybridoma technology. The mAbs were evaluated for their reactivity and neutralization activity. The mAbs were generated through hybridoma technology by the fusion of myeloma cells and spleen cells from MERS-CoV-S1 immunized mice. The resulting hybridomas were screened for antibody production using enzyme-linked immunosorbent assays (ELISA). RESULTS ELISA results demonstrated that all three mAbs exhibited strong reactivity against the MERS-CoV S1-antigen. Similarly, dot-ELISA revealed their ability to specifically recognize viral components, indicating their potential for diagnostic applications. Under non-denaturing conditions, Western blot showed the mAbs to have robust reactivity against a specific band at 116 KDa, corresponding to a putative MERS-CoV S1-antigen. However, no reactive bands were observed under denaturing conditions, suggesting that the antibodies recognize conformational epitopes. The neutralization assay showed no in vitro reactivity against MERS-CoV. CONCLUSION This study successfully generated three mouse monoclonal antibodies against MERS-CoV using hybridoma technology. The antibodies exhibited strong reactivity against MERS-CoV antigens using ELISA and dot ELISA assays. Taken together, these findings highlight the significance of these mAbs for potential use as valuable tools for MERS-CoV research and diagnosis (community and field-based surveillance and viral antigen detection).

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针对中东呼吸综合征冠状病毒尖峰蛋白 S1 亚基的三种新型小鼠单克隆抗体的开发和特征描述。

背景中东呼吸综合征冠状病毒是一种高致病性病毒，对公共卫生构成重大威胁。目的本研究的目的是开发和鉴定针对中东呼吸综合征冠状病毒（MERS-CoV）尖峰蛋白 S1 亚基的新型小鼠单克隆抗体。对这些 mAbs 的反应性和中和活性进行了评估。这些 mAbs 是通过杂交瘤技术将骨髓瘤细胞和 MERS-CoV-S1 免疫小鼠的脾脏细胞融合产生的。结果表明，这三种 mAbs 对 MERS-CoV S1 抗原都有很强的反应性。同样，点式酶联免疫吸附试验（dot-ELISA）也显示了它们特异性识别病毒成分的能力，表明它们具有诊断应用的潜力。在非变性条件下，Western 印迹显示 mAbs 对 116 KDa 的特异性条带有很强的反应性，该条带与推测的 MERS-CoV S1 抗原相对应。然而，在变性条件下没有观察到反应带，这表明抗体能识别构象表位。结论本研究利用杂交瘤技术成功制备了三种针对 MERS-CoV 的小鼠单克隆抗体。通过酶联免疫吸附和点酶联免疫吸附试验，这些抗体对 MERS-CoV 抗原有很强的反应性。综上所述，这些研究结果凸显了这些 mAbs 作为 MERS-CoV 研究和诊断（社区和现场监测以及病毒抗原检测）的重要潜在用途。

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来源期刊

Human Antibodies Medicine-Immunology and Allergy

CiteScore

3.50

自引率

0.00%

发文量

期刊介绍： Human Antibodies is an international journal designed to bring together all aspects of human hybridomas and antibody technology under a single, cohesive theme. This includes fundamental research, applied science and clinical applications. Emphasis in the published articles is on antisera, monoclonal antibodies, fusion partners, EBV transformation, transfections, in vitro immunization, defined antigens, tissue reactivity, scale-up production, chimeric antibodies, autoimmunity, natural antibodies/immune response, anti-idiotypes, and hybridomas secreting interesting growth factors. Immunoregulatory molecules, including T cell hybridomas, will also be featured.