NPJ VaccinesPub Date : 2026-04-18DOI: 10.1038/s41541-026-01411-1
Holly Janes, Youyi Fong, Ying Huang, David Benkeser, Elizabeth J Kelly, Ian Hirsch, Ann Marie Stanley, Tonya Villafana, Christos J Petropoulos, Andrew Leith, Deanne Haugaard, Bill Webb, Yiwen Lu, Chenchen Yu, Bhavesh Borate, Lars W P van der Laan, Nima S Hejazi, April K Randhawa, Michele P Andrasik, James G Kublin, Margaret Brewinski Isaacs, Mamodikoe Makhene, Tina Tong, Merlin L Robb, Lawrence Corey, Kathleen M Neuzil, Dean Follmann, Ann R Falsey, Magdalena E Sobieszczyk, Richard A Koup, Peter B Gilbert
{"title":"Correlates of severe and delta COVID-19 in a phase 3 trial of the AZD1222 vaccine.","authors":"Holly Janes, Youyi Fong, Ying Huang, David Benkeser, Elizabeth J Kelly, Ian Hirsch, Ann Marie Stanley, Tonya Villafana, Christos J Petropoulos, Andrew Leith, Deanne Haugaard, Bill Webb, Yiwen Lu, Chenchen Yu, Bhavesh Borate, Lars W P van der Laan, Nima S Hejazi, April K Randhawa, Michele P Andrasik, James G Kublin, Margaret Brewinski Isaacs, Mamodikoe Makhene, Tina Tong, Merlin L Robb, Lawrence Corey, Kathleen M Neuzil, Dean Follmann, Ann R Falsey, Magdalena E Sobieszczyk, Richard A Koup, Peter B Gilbert","doi":"10.1038/s41541-026-01411-1","DOIUrl":"10.1038/s41541-026-01411-1","url":null,"abstract":"<p><p>In the phase 3 AZD1222 COVID-19 vaccine trial, anti-Spike (vaccine-matched and Delta) binding IgG antibody concentration and neutralizing antibody (nAb) titer (vaccine-matched+D614G and Delta), measured four weeks post-dose two (D57), were assessed as correlates of risk of severe COVID-19 and Delta COVID-19 over ~4 to ~13 months (severe) or ~11 months (Delta) post-D57. Using a case-control design, antibodies were measured in baseline SARS-CoV-2-negative per-protocol ChAdOx1 nCoV-19 recipients (19 severe COVID-19 cases, 57 Delta COVID-19 cases, 111 controls). The hazard ratio (HR) of severe COVID-19 per 10-fold vaccine-matched D57 marker increase was 0.16 (95% CI: 0.05, 0.54; p = 0.004) for Spike IgG and 0.13 (0.03, 0.59; p = 0.009) for nAb titer. D57 Delta antibodies were weak correlates of Delta COVID-19: HR per 10-fold increase 0.70 (0.14, 3.47; p = 0.66) for Delta Spike IgG; 0.46 (0.14, 1.47; p = 0.19) for Delta nAb titer. Binding and nAb levels strongly predicted severe COVID-19, even with antibody waning.</p>","PeriodicalId":19335,"journal":{"name":"NPJ Vaccines","volume":" ","pages":""},"PeriodicalIF":7.2,"publicationDate":"2026-04-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13276171/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147717541","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Rational selection of antigenic targets for Group A Streptococcus vaccine: updates, challenges and opportunities.","authors":"Melaku Ashagrie Belete, Istvan Toth, Rachel Stephenson, Mariusz Skwarczynski","doi":"10.1038/s41541-026-01445-5","DOIUrl":"10.1038/s41541-026-01445-5","url":null,"abstract":"<p><p>Group A Streptococcus (GAS) remains a major global health challenge, causing over half a million deaths annually. Despite nearly a century of research, no licensed vaccine is currently available, due to antigenic diversity, complex host-pathogen interactions, risk of autoimmunity and lack of clear correlates of protection. This review discusses recent advances in GAS vaccine research, highlights key challenges and outlines criteria and emerging opportunities for antigen selection.</p>","PeriodicalId":19335,"journal":{"name":"NPJ Vaccines","volume":" ","pages":""},"PeriodicalIF":7.2,"publicationDate":"2026-04-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13276383/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147717567","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"A poorly reactogenic lipid nanoparticle-mRNA vaccine unveils an innate immune pathway for adverse reactions.","authors":"Tomohiro Takano, Keigo Kumagai, Hitoshi Iuchi, Kazutaka Terahara, Aya Mizuike, Eita Sasaki, Yu Adachi, Ryutaro Kotaki, Saya Moriyama, Shinichiro Ota, Mizuki Fujisawa, Tomoharu Mizukami, Kyoko Saito, Masanori Isogawa, Kohei Soga, Haruyo Nakajima-Adachi, Satoshi Hachimura, Kouji Kobiyama, Ken J Ishii, Michiaki Hamada, Masayoshi Fukasawa, Masaharu Shinkai, Takayuki Matsumura, Yoshimasa Takahashi","doi":"10.1038/s41541-026-01441-9","DOIUrl":"10.1038/s41541-026-01441-9","url":null,"abstract":"<p><p>Lipid nanoparticle (LNP)-mRNA vaccines robustly activate immune responses, contributing to their high efficacy and frequent adverse reactions (ARs). Here, we identified an LNP-mRNA formulation with a more favorable balanced immunogenicity-reactogenicity profile. Immune profiling in a mouse model defined the reactogenic LNP-mRNA vaccine as a potent inducer of HMGB1 release, pro-inflammatory cytokine production, and concurrent neutrophil infiltration. HMGB1 induced TNF-α secretion from monocyte subsets, yet in vivo blockade studies revealed the contribution of multiple cytokines (TNF-α, IL-1, and IL-6) to reactogenicity. Among the reactogenic cytokines, IL-1 was identified as the key mediator of vaccine-induced ARs, but was dispensable for humoral immunity. The clinical relevance was confirmed in a well-controlled vaccine cohort where IL-1 pathway activation correlated with fever severity but not with neutralizing antibody titers. We dissected early innate pathways specifically linked to vaccine reactogenicity, providing a rationale for selectively reducing ARs in next-generation vaccines.</p>","PeriodicalId":19335,"journal":{"name":"NPJ Vaccines","volume":" ","pages":""},"PeriodicalIF":7.2,"publicationDate":"2026-04-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13323985/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147699319","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
NPJ VaccinesPub Date : 2026-04-15DOI: 10.1038/s41541-026-01450-8
Wenying Zhao, Lingjin Sun, Peng Wang, Qian Zhao, Ying Li, Yang Li, Hongyang Shi, Man Xing, Weiqian Dai, Dongming Zhou
{"title":"An mRNA vaccine confers enhanced protection against herpes simplex virus through an IFN-I-dependent pathway.","authors":"Wenying Zhao, Lingjin Sun, Peng Wang, Qian Zhao, Ying Li, Yang Li, Hongyang Shi, Man Xing, Weiqian Dai, Dongming Zhou","doi":"10.1038/s41541-026-01450-8","DOIUrl":"10.1038/s41541-026-01450-8","url":null,"abstract":"<p><p>Herpes simplex virus (HSV) types 1 and 2 cause widespread oral or genital infections, but no prophylactic or therapeutic HSV vaccine has been approved to date. In this study, we developed three mRNA vaccine candidates expressing key viral glycoproteins: monovalent gD2, bivalent gD2-gC1, and bivalent gD2-gE1. We assessed their immunogenicity and protective efficacy in a murine model. All candidates elicited robust humoral and cellular immunity and provided significant protection against intravaginal HSV challenge. Notably, the gD2-gE1 vaccine induced markedly stronger immune responses. Mechanistically, its superior immunoprotective efficacy was associated with the stronger IFN‑I response, which thereby enhanced the adaptive immune response. Collectively, our findings provide a scientific rationale and valuable insights for the future development of HSV vaccines.</p>","PeriodicalId":19335,"journal":{"name":"NPJ Vaccines","volume":" ","pages":""},"PeriodicalIF":7.2,"publicationDate":"2026-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13265912/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147691280","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
NPJ VaccinesPub Date : 2026-04-15DOI: 10.1038/s41541-026-01426-8
Johanne Audouze-Chaud, Ann-Katrin Schlosser, Angelika B Riemer
{"title":"The current landscape of therapeutic vaccination approaches for treatment of HPV-dependent malignancies.","authors":"Johanne Audouze-Chaud, Ann-Katrin Schlosser, Angelika B Riemer","doi":"10.1038/s41541-026-01426-8","DOIUrl":"10.1038/s41541-026-01426-8","url":null,"abstract":"<p><p>This review provides an overview of current therapeutic human papillomavirus (HPV) vaccination approaches. We review important criteria for their development, and present vaccines currently being actively followed in clinical trials, for precursor lesions or established cancers. Subsequently, we list approaches that reached clinical trial stage but were abandoned. Lastly, we discuss promising preclinical studies. We conclude with a brief overview of novel in silico tools developed to enhance vaccine design.</p>","PeriodicalId":19335,"journal":{"name":"NPJ Vaccines","volume":" ","pages":""},"PeriodicalIF":7.2,"publicationDate":"2026-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13103388/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147691251","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
NPJ VaccinesPub Date : 2026-04-14DOI: 10.1038/s41541-026-01442-8
Vincent Dussupt, Jaime L Jensen, Angélica Peña Rosado, Marissa Donofrio, Jill Pflugheber, Letzibeth Mendez-Rivera, Rajeshwer S Sankhala, Wei-Hung Chen, Bonnie M Slike, Annika Schmid, Ursula Tran, Lily Metzger, Caroline E Peterson, Amelia K Pinto, Sandhya Vasan, Natalie D Collins, Aaron Farmer, Nelson L Michael, M Gordon Joyce, James D Brien, Shelly J Krebs
{"title":"Targeting the Zika virus envelope domains I and III as a recombinant vaccine protects mice from lethal challenge.","authors":"Vincent Dussupt, Jaime L Jensen, Angélica Peña Rosado, Marissa Donofrio, Jill Pflugheber, Letzibeth Mendez-Rivera, Rajeshwer S Sankhala, Wei-Hung Chen, Bonnie M Slike, Annika Schmid, Ursula Tran, Lily Metzger, Caroline E Peterson, Amelia K Pinto, Sandhya Vasan, Natalie D Collins, Aaron Farmer, Nelson L Michael, M Gordon Joyce, James D Brien, Shelly J Krebs","doi":"10.1038/s41541-026-01442-8","DOIUrl":"10.1038/s41541-026-01442-8","url":null,"abstract":"<p><p>Zika virus (ZIKV) vaccine candidates developed through Phase I clinical trials are based on the full-length envelope glycoprotein (E), which presents both desirable and undesirable antigenic determinants. Among the latter, the conserved fusion loop epitope (FLE) within domain II is a major target for flavivirus cross-reactive and poorly neutralizing responses. To eliminate unwanted FLE targeting, we redesigned ZIKV E using a reverse vaccinology approach, excising domain II and allowing domains I and III (DI-DIII) to fold into an independent subunit harboring key neutralizing epitopes. Ifnar1<sup>-/-</sup> mice vaccinated with ZIKV DI-DIII elicited high ZIKV neutralizing antibodies and were protected from weight loss and death. In addition, sera from DI-DIII vaccinated mice demonstrated a reduced capacity to enhance DENV 1-4 infection in vitro, compared to mice vaccinated with full-length E. This study identifies DI-DIII as a promising immunogen, focusing antibody responses to protective epitopes on ZIKV and minimizing the elicitation of unwanted responses.</p>","PeriodicalId":19335,"journal":{"name":"NPJ Vaccines","volume":" ","pages":""},"PeriodicalIF":7.2,"publicationDate":"2026-04-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13270120/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147691314","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
NPJ VaccinesPub Date : 2026-04-10DOI: 10.1038/s41541-026-01439-3
Kee Woong Kwon, Hongmin Kim, Hagyu Kim, Roland Brosch, Sung Jae Shin
{"title":"An ESAT-6-convergent prime-boost vaccination combining recombinant BCG expressing Mycobacterium marinum ESX-1 and ESAT-6/GLA-SE improves TB protection.","authors":"Kee Woong Kwon, Hongmin Kim, Hagyu Kim, Roland Brosch, Sung Jae Shin","doi":"10.1038/s41541-026-01439-3","DOIUrl":"10.1038/s41541-026-01439-3","url":null,"abstract":"<p><p>Although Bacille Calmette-Guérin (BCG) protects children against disseminated tuberculosis (TB), its limited efficacy against adult pulmonary TB underscores the need for improved vaccination strategies. We previously developed a recombinant BCG strain expressing the ESX-1 type VII secretion system of Mycobacterium marinum (BCG::ESX-1<sup>Mmar</sup>), which enhances immunogenicity through cytosolic immune signaling while maintaining low virulence. Here, we evaluated an ESAT-6-convergent prime-boost vaccination strategy in which mice were primed with ESX-1-competent BCG::ESX-1<sup>Mmar</sup> and subsequently boosted with Mycobacterium tuberculosis (Mtb)-derived ESAT-6 formulated with the TLR4 agonist adjuvant GLA-SE. Compared with ESAT-6/GLA-SE boosting following parental BCG priming, the BCG::ESX-1<sup>Mmar</sup>-primed regimen robustly increased antigen-specific CD4⁺ T cells localized within the lung parenchyma. This strategy markedly enhanced polyfunctional Th1 responses against ESAT-6 and PPD, exceeding those induced by BCG::ESX-1<sup>Mmar</sup> alone or the conventional BCG-prime/subunit-boost approach. Importantly, ESAT-6 boosting of recombinant BCG::ESX-1<sup>Mmar</sup> conferred superior long-term protection against hypervirulent Mtb challenge and significantly reduced pulmonary inflammation. Together, these findings demonstrate that leveraging an ESX-1-competent recombinant BCG platform for targeted ESAT-6 boosting can overcome key limitations of classical BCG vaccination and represents a promising strategy for next-generation TB immunization.</p>","PeriodicalId":19335,"journal":{"name":"NPJ Vaccines","volume":" ","pages":""},"PeriodicalIF":7.2,"publicationDate":"2026-04-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13253831/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147654732","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
NPJ VaccinesPub Date : 2026-04-10DOI: 10.1038/s41541-026-01437-5
Pauline Saint-Charles, Magdalena Hagen, Gaëlle Autaa, Elske Bijvank, Lisa Beckers, Josine van Beek, Debbie van Baarle, Birgit Weinberger, Victor Appay
{"title":"Influenza vaccine-specific CD4<sup>+</sup> T cell responses are impaired in older adults.","authors":"Pauline Saint-Charles, Magdalena Hagen, Gaëlle Autaa, Elske Bijvank, Lisa Beckers, Josine van Beek, Debbie van Baarle, Birgit Weinberger, Victor Appay","doi":"10.1038/s41541-026-01437-5","DOIUrl":"10.1038/s41541-026-01437-5","url":null,"abstract":"<p><p>Immune defenses decline with age, increasing susceptibility to influenza. Vaccination remains the most effective strategy to prevent severe disease and death, but its efficacy is reduced in older adults, particularly against influenza A(H3N2). The mechanisms underlying this age-related decline in vaccine-specific antibody responses remain unclear. We investigated the magnitude and quality of influenza-specific T-cell responses following quadrivalent inactivated influenza vaccination in adults aged under (n = 100) or over (n = 120) 65 years. Frequencies of T cells specific to influenza A (H1N1, H3N2) and influenza B (Victoria, Yamagata) strains were measured before and after vaccination. Polyfunctionality of vaccine-induced CD4⁺ and CD8⁺ T cells and immune ageing markers were assessed in a subset of responders (n = 34). Older adults exhibited significantly reduced H3N2-specific CD4⁺ T-cell frequencies (P = 0.01) and polyfunctionality (P = 0.04), which correlated with lower H3N2 hemagglutination inhibition antibody titers (r = 0.42, P = 0.008). Cytomegalovirus seropositivity was associated with diminished influenza-specific CD8⁺ T-cell responses in the older age group (P = 0.01). These findings demonstrate quantitative and qualitative deficiencies in influenza-specific memory T cells with ageing, which may contribute to impaired humoral responses, particularly against H3N2. This highlights the need for vaccines that more effectively enhance cellular immunity in older adults, potentially through improved H3N2 antigen design or alternative vaccine platforms.</p>","PeriodicalId":19335,"journal":{"name":"NPJ Vaccines","volume":" ","pages":""},"PeriodicalIF":7.2,"publicationDate":"2026-04-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13270034/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147654780","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
NPJ VaccinesPub Date : 2026-04-09DOI: 10.1038/s41541-026-01434-8
Swap Ghosh, Carrie L Simms, Sage D Rohrer, Carlos H Castaneda, Joseph W Saelens, Kaley Niehaus, Michael J Swyers, Joseph Russo
{"title":"USTAT: Unified Sequential Template Amplification and Transcription-a fully synthetic mRNA manufacturing platform.","authors":"Swap Ghosh, Carrie L Simms, Sage D Rohrer, Carlos H Castaneda, Joseph W Saelens, Kaley Niehaus, Michael J Swyers, Joseph Russo","doi":"10.1038/s41541-026-01434-8","DOIUrl":"10.1038/s41541-026-01434-8","url":null,"abstract":"<p><p>mRNA vaccines have emerged as a transformative modality for infectious disease prevention. In response to the emergence of SARS-CoV-2, large-scale in vitro transcription (IVT) of mRNA vaccines was developed. Large-scale IVT currently relies on linearized plasmid DNA (pDNA) as a template for mRNA production. Linearized pDNA production presents several challenges at manufacturing scale, including removal of residual host-cell DNA, protein, endotoxins, and antibiotics. Additionally, pDNA-derived sequences irrelevant to mRNA production must be removed from the final product. Finally, the generation of linear pDNA template is laborious, which reduces mRNA production speed, a renowned advantage of this technology. Enzymatic DNA amplification strategies such as rolling circle amplification (RCA) of a synthetic circular DNA molecule offer a rapid, isothermal reaction as an alternative to pDNA. Therefore, we have developed a fully synthetic, single-vessel mRNA manufacturing platform. Beginning with a chemically synthesized circular DNA template, we amplify via a fit-for-purpose RCA, linearize with a TypeIIS restriction enzyme (RE), and perform IVT in a single vessel. The entire process-from circular template to mRNA-can be completed in as little as two days. This method, termed Unified Sequential Template Amplification and Transcription (USTAT), eliminates bacterial components, large volume pDNA production, and enables rapid, modular mRNA production.</p>","PeriodicalId":19335,"journal":{"name":"NPJ Vaccines","volume":" ","pages":""},"PeriodicalIF":7.2,"publicationDate":"2026-04-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13246738/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147645935","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
NPJ VaccinesPub Date : 2026-04-08DOI: 10.1038/s41541-026-01435-7
Jorge Armero-Gimenez, Janis Schleusner, Ruud H P Wilbers, Arjen Schots, Livia Spiga, John S Tregoning, Jonathan C Brown, Jie Zhou, Hannes Juergens, Ricarda Finnern, Charles Williams
{"title":"Noncanonical amino acids enable plug and play vaccine platform in the ALiCE cell free system.","authors":"Jorge Armero-Gimenez, Janis Schleusner, Ruud H P Wilbers, Arjen Schots, Livia Spiga, John S Tregoning, Jonathan C Brown, Jie Zhou, Hannes Juergens, Ricarda Finnern, Charles Williams","doi":"10.1038/s41541-026-01435-7","DOIUrl":"10.1038/s41541-026-01435-7","url":null,"abstract":"<p><p>Non-canonical amino acids (ncaas) are increasingly used in vaccinology to improve vaccine adaptability and immunogenicity. Cell-free protein synthesis (CFPS) offers a promising route for site-specific ncaa incorporation, but conventional prokaryotic CFPS systems show limitations to produce complex proteins requiring post-translational modifications while eukaryotic systems have historically been difficult to scale and show low protein yields. Here we establish efficient site-specific introduction of ncaas into complex proteins with the high-yielding and scalable eukaryotic tobacco BY-2 CFPS system (BYL), commercialized as ALiCE®. ncaa incorporation yields reached up to 2 mg/ml with linear scalability up to 10 ml. We applied ncaa incorporation in BYL to enable click chemistry bioconjugation of the receptor binding domain (RBD) of influenza hemagglutinin to pre-assembled hepatitis B core (HBc) virus-like particles (VLPs). The resulting VLP-RBD conjugates exhibited hemagglutination activity, unlike the individual components, and protected mice from weight loss after influenza challenge. This research thus enables ncaa introduction for recombinant proteins produced in BYL, constructing a novel plug-and-play vaccine platform and further expanding the capabilities of BYL to produce vaccine candidates and other proteins of interest.</p>","PeriodicalId":19335,"journal":{"name":"NPJ Vaccines","volume":" ","pages":""},"PeriodicalIF":7.2,"publicationDate":"2026-04-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13068933/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147639388","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}