{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,23]],"date-time":"2026-09-23T22:18:54Z","timestamp":1790201934155,"version":"4.1.0"},"reference-count":19,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2026,7,3]],"date-time":"2026-07-03T00:00:00Z","timestamp":1783036800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100016302","name":"Coalition for Epidemic Preparedness Innovations","doi-asserted-by":"publisher","award":["276869"],"award-info":[{"award-number":["276869"]}],"id":[{"id":"10.13039\/100016302","id-type":"DOI","asserted-by":"publisher"}]},{"award":["276869"],"award-info":[{"award-number":["276869"]}],"id":[{"id":"https:\/\/ror.org\/01t08q012","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Vaccines"],"abstract":"<jats:p>Background\/Objectives: Vesicular Stomatitis Virus (VSV)-vectored vaccines represent a versatile platform for the development of vaccines against infectious diseases. Replication-competent recombinant VSV vectors in which the native glycoprotein (G) is replaced with a heterologous antigen (rVSV\u0394G) are widely used and have demonstrated clinical success. In addition to antigen-specific responses, these vaccines induce humoral immunity directed against vector components, which primarily reflects vector exposure and contributes to the overall characterization of vaccine-induced immunity. Standardized assays for quantifying such vector-directed responses are therefore of increasing importance. Methods: We developed and qualified a quantitative enzyme-linked immunosorbent assay (ELISA) for the detection of human IgG antibodies against VSV-Nucleoprotein (N). Assay development included optimization of antigen coating, blocking conditions, and detection reagents. A 10-point standard curve was established using pooled human serum, and assay performance was evaluated by assessing dynamic range, sensitivity, cut point, dilutional linearity, precision, robustness, and sample stability. Results: The optimized assay utilized a coating concentration of 2 \u03bcg\/mL VSV-N antigen (100 ng\/well) and 1% casein as the blocking buffer. The assay demonstrated a dynamic range of 0.33\u201341.66 arbitrary units per milliliter (AU\/mL) with excellent curve fit (R2 &gt; 0.996). The cut point was established at an OD450 of 0.286. Precision across intra-assay, inter-assay, and inter-operator evaluations met predefined acceptance criteria (\u226425% CV). The assay was robust across different plate washers and readers and maintained performance following up to three freeze\u2013thaw cycles. Conclusions: This study describes a robust and reproducible ELISA for quantifying anti-VSV-N IgG responses. The assay provides a fit-for-purpose tool for assessing vector-directed humoral immunity and supports standardized immunogenicity evaluations across VSV-vectored vaccine studies.<\/jats:p>","DOI":"10.3390\/vaccines14070592","type":"journal-article","created":{"date-parts":[[2026,7,3]],"date-time":"2026-07-03T07:59:47Z","timestamp":1783065587000},"page":"592","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Development and Qualification of a VSV-N IgG ELISA to Assess Vector-Directed Humoral Immunity in VSV-Vectored Vaccine Studies"],"prefix":"10.3390","volume":"14","author":[{"given":"Morolayo","family":"Ayorinde","sequence":"first","affiliation":[{"name":"IAVI Human Immunology Laboratory, Imperial College London, London SW10 9NH, UK"},{"name":"IAVI, New York, NY 10004, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Claire","family":"Streatfield","sequence":"additional","affiliation":[{"name":"IAVI Human Immunology Laboratory, Imperial College London, London SW10 9NH, UK"},{"name":"IAVI, New York, NY 10004, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vanaja","family":"Kakarla","sequence":"additional","affiliation":[{"name":"IAVI Human Immunology Laboratory, Imperial College London, London SW10 9NH, UK"},{"name":"IAVI, New York, NY 10004, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Faith","family":"Sigei","sequence":"additional","affiliation":[{"name":"IAVI Human Immunology Laboratory, Imperial College London, London SW10 9NH, UK"},{"name":"IAVI, New York, NY 10004, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rachel","family":"Bromell","sequence":"additional","affiliation":[{"name":"IAVI Human Immunology Laboratory, Imperial College London, London SW10 9NH, UK"},{"name":"IAVI, New York, NY 10004, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Arianna","family":"Marini","sequence":"additional","affiliation":[{"name":"IAVI Human Immunology Laboratory, Imperial College London, London SW10 9NH, UK"},{"name":"IAVI, New York, NY 10004, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0569-6049","authenticated-orcid":false,"given":"Marija","family":"Zaric","sequence":"additional","affiliation":[{"name":"IAVI Human Immunology Laboratory, Imperial College London, London SW10 9NH, UK"},{"name":"IAVI, New York, NY 10004, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,7,3]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1111\/imm.12829","article-title":"Novel viral vectors in infectious diseases","volume":"153","author":"Humphreys","year":"2018","journal-title":"Immunology"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"149","DOI":"10.1038\/s41392-023-01408-5","article-title":"Viral vectored vaccines: Design, development, preventive and therapeutic applications in human diseases","volume":"8","author":"Wang","year":"2023","journal-title":"Signal Transduct. 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Hyg."}],"container-title":["Vaccines"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-393X\/14\/7\/592\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,3]],"date-time":"2026-07-03T09:17:59Z","timestamp":1783070279000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-393X\/14\/7\/592"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,3]]},"references-count":19,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2026,7]]}},"alternative-id":["vaccines14070592"],"URL":"https:\/\/doi.org\/10.3390\/vaccines14070592","relation":{},"ISSN":["2076-393X"],"issn-type":[{"value":"2076-393X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,3]]}}}