Highly scalable prefusion-stabilized RSV F vaccine with enhanced immunogenicity and robust protection

Abstract Stabilizing the RSV F protein in its prefusion conformation is crucial for effective vaccine development but has remained a significant challenge. Traditional stabilization methods, such as disulfide bonds and cavity-filling mutations, have been labor-intensive and have often resulted in su...

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Main Authors: Qingrui Huang, Qingyun Lang, Yao Li, Fengjie Wang, Xiaonan Han, Ling Luo, Xiaomin Duan, Xuerong Cao, Yue Wang, Yu Bai, Yaxin Song, Yanpeng Xu, Lianlian Bian, Chenyan Gao, Linqing Zhao, Jinghua Yan
Format: Article
Language:English
Published: Nature Portfolio 2025-08-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-63084-z
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author Qingrui Huang
Qingyun Lang
Yao Li
Fengjie Wang
Xiaonan Han
Ling Luo
Xiaomin Duan
Xuerong Cao
Yue Wang
Yu Bai
Yaxin Song
Yanpeng Xu
Lianlian Bian
Chenyan Gao
Linqing Zhao
Jinghua Yan
author_facet Qingrui Huang
Qingyun Lang
Yao Li
Fengjie Wang
Xiaonan Han
Ling Luo
Xiaomin Duan
Xuerong Cao
Yue Wang
Yu Bai
Yaxin Song
Yanpeng Xu
Lianlian Bian
Chenyan Gao
Linqing Zhao
Jinghua Yan
author_sort Qingrui Huang
collection DOAJ
description Abstract Stabilizing the RSV F protein in its prefusion conformation is crucial for effective vaccine development but has remained a significant challenge. Traditional stabilization methods, such as disulfide bonds and cavity-filling mutations, have been labor-intensive and have often resulted in suboptimal expression levels. Here, we report the design of an RSV prefusion F (preF) antigen using a proline-scanning strategy, incorporating seven proline substitutions to achieve stabilization. The resulting variant, preF7P, is structurally and biochemically validated to maintain the correct prefusion state. PreF7P demonstrates superior immunogenicity with a 1.8-fold increase in neutralizing antibody titers when compared to DS-cav2, and provides protection from clinical disease against both RSV A and B strains in female murine and female cotton rat models. In clinical development, preF7P exhibits high expression levels (~10 g/L) in clinical-grade CHO cells. The clinical-grade vaccine elicits robust immunogenic responses across female mice, female SD rats, and both male and female cynomolgus macaques, significantly boosting RSV pre-infection neutralizing antibody titers, and providing sustained protection for at least six months in female mice. This proline-scanning strategy offers a streamlined approach for stabilizing class I fusion proteins, potentially accelerating the development of vaccines for other pathogens.
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spelling doaj-art-de4949e7da5f443082b4919fa5c97dd52025-08-24T11:38:32ZengNature PortfolioNature Communications2041-17232025-08-0116111510.1038/s41467-025-63084-zHighly scalable prefusion-stabilized RSV F vaccine with enhanced immunogenicity and robust protectionQingrui Huang0Qingyun Lang1Yao Li2Fengjie Wang3Xiaonan Han4Ling Luo5Xiaomin Duan6Xuerong Cao7Yue Wang8Yu Bai9Yaxin Song10Yanpeng Xu11Lianlian Bian12Chenyan Gao13Linqing Zhao14Jinghua Yan15Changping LaboratoryChangping LaboratoryChangping LaboratoryLaboratory of Virology, Beijing Key Laboratory of Etiology of Viral Diseases in Children, Capital Institute of PediatricsCAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of SciencesChangping LaboratoryChangping LaboratoryChangping LaboratoryCAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of SciencesChangping LaboratoryChangping LaboratoryLaboratory of Virology, Beijing Key Laboratory of Etiology of Viral Diseases in Children, Capital Institute of PediatricsChangping LaboratoryChangping LaboratoryLaboratory of Virology, Beijing Key Laboratory of Etiology of Viral Diseases in Children, Capital Institute of PediatricsChangping LaboratoryAbstract Stabilizing the RSV F protein in its prefusion conformation is crucial for effective vaccine development but has remained a significant challenge. Traditional stabilization methods, such as disulfide bonds and cavity-filling mutations, have been labor-intensive and have often resulted in suboptimal expression levels. Here, we report the design of an RSV prefusion F (preF) antigen using a proline-scanning strategy, incorporating seven proline substitutions to achieve stabilization. The resulting variant, preF7P, is structurally and biochemically validated to maintain the correct prefusion state. PreF7P demonstrates superior immunogenicity with a 1.8-fold increase in neutralizing antibody titers when compared to DS-cav2, and provides protection from clinical disease against both RSV A and B strains in female murine and female cotton rat models. In clinical development, preF7P exhibits high expression levels (~10 g/L) in clinical-grade CHO cells. The clinical-grade vaccine elicits robust immunogenic responses across female mice, female SD rats, and both male and female cynomolgus macaques, significantly boosting RSV pre-infection neutralizing antibody titers, and providing sustained protection for at least six months in female mice. This proline-scanning strategy offers a streamlined approach for stabilizing class I fusion proteins, potentially accelerating the development of vaccines for other pathogens.https://doi.org/10.1038/s41467-025-63084-z
spellingShingle Qingrui Huang
Qingyun Lang
Yao Li
Fengjie Wang
Xiaonan Han
Ling Luo
Xiaomin Duan
Xuerong Cao
Yue Wang
Yu Bai
Yaxin Song
Yanpeng Xu
Lianlian Bian
Chenyan Gao
Linqing Zhao
Jinghua Yan
Highly scalable prefusion-stabilized RSV F vaccine with enhanced immunogenicity and robust protection
Nature Communications
title Highly scalable prefusion-stabilized RSV F vaccine with enhanced immunogenicity and robust protection
title_full Highly scalable prefusion-stabilized RSV F vaccine with enhanced immunogenicity and robust protection
title_fullStr Highly scalable prefusion-stabilized RSV F vaccine with enhanced immunogenicity and robust protection
title_full_unstemmed Highly scalable prefusion-stabilized RSV F vaccine with enhanced immunogenicity and robust protection
title_short Highly scalable prefusion-stabilized RSV F vaccine with enhanced immunogenicity and robust protection
title_sort highly scalable prefusion stabilized rsv f vaccine with enhanced immunogenicity and robust protection
url https://doi.org/10.1038/s41467-025-63084-z
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