Ingrid Sander, Sabine Kespohl, Eva Zahradnik, Philipp G?cke, Ingolf Hosbach, Burkhard L Herrmann, Thomas Brüning, Monika Raulf
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引用次数: 3
摘要
目的对SARS-CoV-2抗原的体液免疫反应进行标准化定量分析，有助于估计免疫的程度和持续时间。目的是开发酶联免疫吸附测定法(elisa)，用于定量人抗SARS-CoV-2抗原的IgG抗体。方法建立针对人IgG和重组SARS-CoV-2抗原(Spike-S1和Nucleocapsid)的单克隆抗体的酶联免疫吸附试验。采用WHO 67/086免疫球蛋白和WHO 20/136 SARS-CoV-2参比进行标准化。对新冠病毒阳性受试者(144例)、大流行前对照组(135例)和BioNTech-Pfizer BNT162b2疫苗接种个体(48例)的血清进行分析。研究组的血清还使用赛默飞世尔公司的euroimmune sars - cov -2 elisa和定量s1特异性荧光酶免疫分析法(FEIA)进行检测。结果ELISA结果与世界卫生组织的参考文献一致，具有可重复性和可追溯性。在研究组中，抗s1 - igg的中位浓度为102 BAU mL - 1，而接种组在第一次和第二次接种后分别为100和1457 BAU mL - 1。在受试者工作特征(ROC)分析中，elisa的曲线下面积(AUC)分别为0.965 (S1)和0.955 (Nucleocapsid)，特异性分别为1 (S1)和0.963 (Nucleocapsid)，灵敏度分别为0.903 (S1)和0.833 (Nucleocapsid)。相比之下，商业检测方法(S1- feia, S1和核衣壳ELISA EuroImmun)在研究组的灵敏度分别为0.764,0.875和0.882。结论elisa定量检测IgG与SARS-CoV-2抗原结合具有良好的分析和临床性能特点，单位可溯源至国际标准。
Quantitative measurement of IgG to SARS-CoV-2 antigens using monoclonal antibody-based enzyme-linked immunosorbent assays
Objective
Standardised quantitative analysis of the humoral immune response to SARS-CoV-2 antigens may be useful for estimating the extent and duration of immunity. The aim was to develop enzyme-linked immunosorbent assays (ELISAs) for the quantification of human IgG antibodies against SARS-CoV-2 antigens.
Methods
Enzyme-linked immunosorbent assays were developed based on monoclonal antibodies against human IgG and recombinant SARS-CoV-2 antigens (Spike-S1 and Nucleocapsid). The WHO 67/086 immunoglobulin and WHO 20/136 SARS-CoV-2 references were used for standardisation. Sera of a study group of COVID-19-positive subjects (n = 144), pre-pandemic controls (n = 135) and individuals vaccinated with BioNTech–Pfizer BNT162b2 vaccine (n = 48) were analysed. The study group sera were also tested using EuroImmun SARS-CoV-2-ELISAs and a quantitative S1-specific fluorescence enzyme immunoassay (FEIA) from Thermo Fisher.
Results
The ELISA results were repeatable and traceable to international units because of their parallelism to both WHO references. In the study group, median anti-S1-IgG concentrations were 102 BAU mL−1, compared to 100 and 1457 BAU mL−1 in the vaccination group after first and second vaccination, respectively. The ELISAs achieved an area under the curve (AUC) of 0.965 (S1) and 0.955 (Nucleocapsid) in receiver operating characteristic (ROC) analysis, and a specificity of 1 (S1) and 0.963 (Nucleocapsid) and sensitivity of 0.903 (S1) and 0.833 (Nucleocapsid) at the maximum Youden index. In comparison, the commercial assays (S1-FEIA, S1 and Nucleocapsid ELISA EuroImmun) achieved sensitivities of 0.764, 0.875 and 0.882 in the study group, respectively.
Conclusions
The quantitative ELISAs to measure IgG binding to SARS-CoV-2 antigens have good analytical and clinical performance characteristics and units traceable to international standards.
期刊介绍：
Clinical & Translational Immunology is an open access, fully peer-reviewed journal devoted to publishing cutting-edge advances in biomedical research for scientists and physicians. The Journal covers fields including cancer biology, cardiovascular research, gene therapy, immunology, vaccine development and disease pathogenesis and therapy at the earliest phases of investigation.
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