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利用重组植物生产蛋白开发新冠病毒中和抗体检测方法。

Development of SARS-CoV-2 neutralizing antibody detection assay by using recombinant plant-produced proteins.

作者信息

Jirarojwattana Perawat, Shanmugaraj Balamurugan, Rattanapisit Kaewta, Phoolcharoen Waranyoo

机构信息

Center of Excellence in Plant-produced Pharmaceuticals, Chulalongkorn University, Bangkok 10330, Thailand.

Department of Pharmacognosy and Pharmaceutical Botany, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok 10330, Thailand.

出版信息

Biotechnol Rep (Amst). 2023 Jun;38:e00796. doi: 10.1016/j.btre.2023.e00796. Epub 2023 Apr 6.

Abstract

Detecting immunity against SARS-CoV-2 is vital for evaluating vaccine response and natural infection, but conventional virus neutralization test (cVNT) requires BSL3 and live viruses, and pseudo-virus neutralization test (pVNT) needs specialized equipment and trained professionals. The surrogate virus neutralization test (sVNT) was developed to overcome these limitations. This study explored the use of angiotensin converting enzyme 2 (ACE2) produced from for the development of an affordable neutralizing antibodies detection assay. The results showed that the plant-produced ACE2 can bind to the receptor binding domain (RBD) of the SARS-CoV-2, and was used to develop sVNT with plant-produced RBD protein. The sVNT developed using plant-produced proteins showed high sensitivity and specificity when validated with a group of 30 RBD vaccinated mice sera and the results were correlated with cVNT titer. This preliminary finding suggests that the plants could offer a cost-effective platform for producing diagnostic reagents.

摘要

检测针对严重急性呼吸综合征冠状病毒2(SARS-CoV-2)的免疫力对于评估疫苗反应和自然感染至关重要,但传统病毒中和试验(cVNT)需要生物安全三级实验室和活病毒,而假病毒中和试验(pVNT)需要专门设备和经过培训的专业人员。为克服这些局限性而开发了替代病毒中和试验(sVNT)。本研究探索了利用植物产生的血管紧张素转换酶2(ACE2)来开发一种经济实惠的中和抗体检测方法。结果表明,植物产生的ACE2可与SARS-CoV-2的受体结合域(RBD)结合,并用于与植物产生的RBD蛋白一起开发sVNT。当用一组30只接种RBD疫苗的小鼠血清进行验证时,利用植物产生的蛋白质开发的sVNT显示出高灵敏度和特异性,且结果与cVNT滴度相关。这一初步发现表明,植物可为生产诊断试剂提供一个具有成本效益的平台。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0473/10130755/c0efed5f252d/gr1.jpg

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