Robust validation and performance comparison of immunogenicity assays assessing IgG and neutralizing antibodies to SARS-CoV-2.
Robust validation and performance comparison of immunogenicity assays assessing IgG and neutralizing antibodies to SARS-CoV-2.
复制标题
DOI：
10.1371/journal.pone.0262922
复制
发表时间：
2022
期刊：
影响因子：
3.7
通讯作者：
Kierstead L
中科院分区：
文献类型：
--
作者：
Bonhomme ME;Bonhomme CJ;Strelow L;Chaudhari A;Howlett A;Breidenbach C;Hester J;Hammond C;Fuzy M;Harvey L;Swanner V;Ellis J;Greway RR;Pisciella VA;Green T;Kierstead L
为了对候选的SARS-CoV-2疫苗之间的免疫原性进行基准测试，需要标准化的、经过验证的免疫原性测定。在本文中，我们报告了用于验证免疫原性分析的设计和标准，以及用于评估人类血清中功能性免疫反应和抗体滴度的血清学和功能分析的验证结果。利用参考标准的定量细胞微中和（MNT）方法检测人血清中抗SARS-CoV-2刺突蛋白中和抗体，Meso Scale Discovery的多重电化学发光（MSD ECL）方法检测针对SARS-CoV-2刺突、核衣壳和受体结合域（RBD）蛋白的免疫球蛋白G （IgG）抗体，评估其精密度、准确性、稀释线性、选择性、使用2019冠状病毒病（COVID-19）确诊的康复供体汇集的人类血清进行特异性分析。两种检测方法均满足预先设定的精密度、相对准确度、稀释线性、选择性和特异性的接受标准。两种检测方法均显示对不同的SARS-CoV-2抗原或检测病毒具有高特异性，且与季节性冠状病毒无显著交叉反应性。一项比较MNT法中和活性与MSD ECL法测定的IgG的评估显示，在恢复期和接种者的血清中，中和活性与抗刺突和RBD蛋白的抗体数量之间存在很强的相关性。最后，根据世卫组织参考标准对MNT测定法进行了校准，以便能够在国际单位报告结果，从而便于比较不同测定法和/或实验室产生的免疫原性数据。MSD ECL试验之前已进行过校准。总之，这些经过验证的评估SARS-CoV-2疫苗接种后功能性免疫反应和抗体滴度的方法可以为准确比较不同疫苗和/或疫苗接种方案的免疫反应提供一种相对简单的标准化方法。
To enable benchmarking of immunogenicity between candidate severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccines, there is a need for standardized, validated immunogenicity assays. In this article, we report the design and criteria used to validate immunogenicity assays and the outcome of the validation of serologic and functional assays for the evaluation of functional immune response and antibody titers in human serum. A quantitative cell-based microneutralization (MNT) assay, utilizing a reference standard, for detecting anti-SARS-CoV-2 spike protein-neutralizing antibodies in human serum and Meso Scale Discovery’s multiplex electrochemiluminescence (MSD ECL) assay for immunoglobulin G (IgG) antibodies to SARS-CoV-2 spike, nucleocapsid, and receptor-binding domain (RBD) proteins were assessed for precision, accuracy, dilutional linearity, selectivity, and specificity using pooled human serum from coronavirus disease 2019 (COVID-19)-confirmed recovered donors. Both assays met prespecified acceptance criteria for precision, relative accuracy, dilutional linearity, selectivity, and specificity. Both assays demonstrated high specificity for the different SARS-CoV-2 antigens or virus tested, and no significant cross-reactivity with seasonal coronaviruses. An evaluation to compare the neutralizing activity in the MNT assay to the IgG measured using the MSD ECL assay showed a strong correlation between the presence of neutralizing activity and amount of antibodies against the spike and RBD proteins in sera from both convalescent and vaccinated individuals. Finally, the MNT assay was calibrated to the WHO reference standard to enable reporting of results in international units, thus facilitating comparison of immunogenicity data generated by different assays and/or laboratories. The MSD ECL assay has previously been calibrated. In conclusion, these validated assays for the evaluation of functional immune response and antibody titers following SARS-CoV-2 vaccination could provide a relatively simple standardized approach for accurately comparing immune responses to different vaccines and/or vaccination regimens.
登录
查看更多内容
DOI：
10.1038/s41577-020-00455-1
发表时间：
2020-11
期刊：
Nature reviews. Immunology
影响因子：
--
作者：
Parker EPK;Shrotri M;Kampmann B
通讯作者：
Kampmann B
影响因子：
64.8
作者：
Mulligan, Mark J.;Lyke, Kirsten E.;Jansen, Kathrin U.
通讯作者：
Jansen, Kathrin U.
影响因子：
64.5
作者：
Korber, Bette;Fischer, Will M.;Montefiori, David C.
通讯作者：
Montefiori, David C.
影响因子：
168.9
作者：
Folegatti, Pedro M.;Ewer, Katie J.;Pollard, Andrew J.
通讯作者：
Pollard, Andrew J.
影响因子：
5.5
作者：
Chu L;McPhee R;Huang W;Bennett H;Pajon R;Nestorova B;Leav B;mRNA-1273 Study Group
通讯作者：
mRNA-1273 Study Group