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利用新冠病毒三聚体刺突蛋白的新型竞争性酶联免疫吸附测定法可识别混合血清中超过RBD-RBM特异性中和抗体。

Novel Competitive ELISA Utilizing Trimeric Spike Protein of SARS-CoV-2, Could Identify More Than RBD-RBM Specific Neutralizing Antibodies in Hybrid Sera.

作者信息

Eliadis Petros, Mais Annie, Papazisis Alexandros, Loxa Eleni K, Dimitriadis Alexios, Sarrigeorgiou Ioannis, Backovic Marija, Agallou Maria, Zouridakis Marios, Karagouni Evdokia, Lazaridis Konstantinos, Mamalaki Avgi, Lymberi Peggy

机构信息

Immunology Laboratory, Immunology Department, Hellenic Pasteur Institute, 11521 Athens, Greece.

Biotechnology Unit, Hellenic Pasteur Institute, 11521 Athens, Greece.

出版信息

Vaccines (Basel). 2024 Aug 13;12(8):914. doi: 10.3390/vaccines12080914.

DOI:10.3390/vaccines12080914
PMID:39204038
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11359269/
Abstract

Since the initiation of the COVID-19 pandemic, there has been a need for the development of diagnostic methods to determine the factors implicated in mounting an immune response against the virus. The most promising indicator has been suggested to be neutralizing antibodies (nAbs), which mainly block the interaction between the Spike protein (S) of SARS-CoV-2 and the host entry receptor ACE2. In this study, we aimed to develop and optimize conditions of a competitive ELISA to measure serum neutralizing titer, using a recombinant trimeric Spike protein modified to have six additional proline residues (S(6P)-HexaPro) and h-ACE2. The results of our surrogate Virus Neutralizing Assay (sVNA) were compared against the commercial sVNT (cPass, Nanjing GenScript Biotech Co., Nanjing City, China), using serially diluted sera from vaccinees, and a high correlation of ID titer values was observed between the two assays. Interestingly, when we tested and compared the neutralizing activity of sera from eleven fully vaccinated individuals who subsequently contracted COVID-19 (hybrid sera), we recorded a moderate correlation between the two assays, while higher sera neutralizing titers were measured with sVNA. Our data indicated that the sVNA, as a more biologically relevant model assay that paired the trimeric S(6P) with ACE2, instead of the isolated RBD-ACE2 pairing cPass test, could identify nAbs other than the RBD-RBM specific ones.

摘要

自新冠疫情爆发以来，一直需要开发诊断方法来确定引发针对该病毒免疫反应的相关因素。最有前景的指标被认为是中和抗体（nAbs），其主要阻断严重急性呼吸综合征冠状病毒2（SARS-CoV-2）的刺突蛋白（S）与宿主进入受体血管紧张素转换酶2（ACE2）之间的相互作用。在本研究中，我们旨在开发并优化一种竞争性酶联免疫吸附测定（ELISA）的条件，以使用经修饰带有六个额外脯氨酸残基的重组三聚体刺突蛋白（S(6P)-HexaPro）和人ACE2来测量血清中和滴度。我们将替代病毒中和试验（sVNA）的结果与商业sVNT（cPass，南京金斯瑞生物科技有限公司，中国南京市）进行比较，使用来自疫苗接种者的系列稀释血清，并且在两种试验之间观察到ID滴度值的高度相关性。有趣的是，当我们测试并比较来自11名随后感染新冠病毒的完全接种疫苗个体（混合血清）的血清中和活性时，我们在两种试验之间记录到中等相关性，而用sVNA测量到更高的血清中和滴度。我们的数据表明，sVNA作为一种更具生物学相关性的模型试验，将三聚体S(6P)与ACE2配对，而不是孤立的受体结合域-血管紧张素转换酶2（RBD-ACE2）配对的cPass试验，可以识别除RBD-RBM特异性中和抗体之外的其他中和抗体。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/51f9/11359269/324d12dce6c1/vaccines-12-00914-g009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/51f9/11359269/b2e7e9e3f5a7/vaccines-12-00914-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/51f9/11359269/9b0135cfe8aa/vaccines-12-00914-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/51f9/11359269/11b51ed42715/vaccines-12-00914-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/51f9/11359269/db60d7a25ae0/vaccines-12-00914-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/51f9/11359269/8a3d6f14b232/vaccines-12-00914-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/51f9/11359269/9a50e7b74a0b/vaccines-12-00914-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/51f9/11359269/04b281aafc97/vaccines-12-00914-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/51f9/11359269/2b953c5522c7/vaccines-12-00914-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/51f9/11359269/324d12dce6c1/vaccines-12-00914-g009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/51f9/11359269/b2e7e9e3f5a7/vaccines-12-00914-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/51f9/11359269/9b0135cfe8aa/vaccines-12-00914-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/51f9/11359269/11b51ed42715/vaccines-12-00914-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/51f9/11359269/db60d7a25ae0/vaccines-12-00914-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/51f9/11359269/8a3d6f14b232/vaccines-12-00914-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/51f9/11359269/9a50e7b74a0b/vaccines-12-00914-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/51f9/11359269/04b281aafc97/vaccines-12-00914-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/51f9/11359269/2b953c5522c7/vaccines-12-00914-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/51f9/11359269/324d12dce6c1/vaccines-12-00914-g009.jpg

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