Validation of a Harmonized Enzyme-Linked-Lectin-Assay (ELLA-NI) Based Neuraminidase Inhibition Assay Standard Operating Procedure (SOP) for Quantification of N1 Influenza Antibodies and the Use of a Calibrator to Improve the Reproducibility of the ELLA-NI With Reverse Genetics Viral and Recombinant Neuraminidase Antigens: A FLUCOP Collaborative Study

Frontiers in Immunology

Serge van de Witte¹  Peter van Amsterdam¹  Emanuele Montomoli²  Claudia Maria Trombetta²  Serena Marchi²  Sylvie Commandeur³  Marie-Clotilde Bernard³  Thierry Ollinger⁴  Sophie Germain⁴  Damien Friel⁴  Rebecca J. Cox⁵  Fan Zhou⁵  Othmar G. Engelhardt⁶  Joanna Waldock⁶  Jerry P. Weir⁷  Vladimir Lugovtsev⁷  Ralf Wagner⁸  Lea Strauß⁸  Hanna Sediri-Schön⁸  Katja Hoschler⁹  Sammy Ho⁹ 

[1] Abbott Healthcare Products B.V., Weesp, Netherlands;Department of Molecular and Developmental Medicine, University of Siena, Siena, Italy;Department of Research and Development, Sanofi Pasteur, Marcy L’Etoile, France;GlaxoSmithKline (GSK), Wavre, Belgium;Influenza Centre, Department of Clinical Sciences, University of Bergen, Bergen, Norway;Influenza Resource Centre, National Institute for Biological Standards and Control, Potters Bar, United Kingdom;Laboratory of DNA Viruses, Division of Viral Products, Office of Vaccines Research and Review, Center for Biologics Evaluation and Research, Food and Drug Administration, Silver Spring, MD, United States;Section viral vaccines, Virology Division, Paul-Ehrlich-Institut, Federal Institute for Vaccines and Biomedicines, Langen, Germany;UK Health Security Agency, Colindale, United Kingdom;

关键词: influenza;    enzyme-linked lectin assay (ELLA);    neuraminidase inhibition (NI) assay;    serology;    standardization;   

DOI  :  10.3389/fimmu.2022.909297

来源: DOAJ

【 摘 要 】

Current vaccination strategies against influenza focus on generating an antibody response against the viral haemagglutination surface protein, however there is increasing interest in neuraminidase (NA) as a target for vaccine development. A critical tool for development of vaccines that target NA or include an NA component is available validated serology assays for quantifying anti-NA antibodies. Additionally serology assays have a critical role in defining correlates of protection in vaccine development and licensure. Standardisation of these assays is important for consistent and accurate results. In this study we first validated a harmonized enzyme-linked lectin assay (ELLA)- Neuraminidase Inhibition (NI) SOP for N1 influenza antigen and demonstrated the assay was precise, linear, specific and robust within classical acceptance criteria for neutralization assays for vaccine testing. Secondly we tested this SOP with NA from influenza B viruses and showed the assay performed consistently with both influenza A and B antigens. Third, we demonstrated that recombinant NA (rNA) could be used as a source of antigen in ELLA-NI. In addition to validating a harmonized SOP we finally demonstrated a clear improvement in inter-laboratory agreement across several studies by using a calibrator. Importantly we showed that the use of a calibrator significantly improved agreement when using different sources of antigen in ELLA-NI, namely reverse genetics viruses and recombinant NA. We provide a freely available and detailed harmonized SOP for ELLA-NI. Our results add to the growing body of evidence in support of developing biological standards for influenza serology.

【 授权许可】

Unknown