{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,16]],"date-time":"2026-06-16T04:36:08Z","timestamp":1781584568129,"version":"3.54.5"},"reference-count":51,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2024,7,30]],"date-time":"2024-07-30T00:00:00Z","timestamp":1722297600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100009708","name":"Novo Nordisk Fonden","doi-asserted-by":"publisher","award":["NNF170C0026778"],"award-info":[{"award-number":["NNF170C0026778"]}],"id":[{"id":"10.13039\/501100009708","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Vaccines"],"abstract":"<jats:p>A vaccine protecting against malaria caused by Plasmodium falciparum is urgently needed. The blood-stage invasion complex PCRCR consists of the five malarial proteins PfPTRAMP, PfCSS, PfRipr, PfCyRPA, and PfRH5. As each subcomponent represents an essential and highly conserved antigen, PCRCR is considered a promising vaccine target. Furthermore, antibodies targeting the complex can block red blood cell invasion by the malaria parasite. However, extremely high titers of neutralizing antibodies are needed for this invasion-blocking effect, and a vaccine based on soluble PfRH5 protein has proven insufficient in inducing a protective response in a clinical trial. Here, we present the results of two approaches to increase the neutralizing antibody titers: (A) immunofocusing and (B) increasing the immunogenicity of the antigen via multivalent display on capsid virus-like particles (cVLPs). The immunofocusing strategies included vaccinating with peptides capable of binding the invasion-blocking anti-PfCyRPA monoclonal antibody CyP1.9, as well as removing non-neutralizing epitopes of PfCyRPA through truncation. Vaccination with PfCyRPA coupled to the AP205 cVLP induced nearly two-fold higher IgG responses compared to vaccinating with soluble PfCyRPA protein. Immunofocusing using a linear peptide greatly increased the neutralizing capacity of the anti-PfCyRPA antibodies. However, significantly lower total anti-PfCyRPA titers were achieved using this strategy. Our results underline the potential of a cVLP-based malaria vaccine including full-length PfCyRPA, which could be combined with other leading malaria vaccine antigens presented on cVLPs.<\/jats:p>","DOI":"10.3390\/vaccines12080859","type":"journal-article","created":{"date-parts":[[2024,7,31]],"date-time":"2024-07-31T10:34:43Z","timestamp":1722422083000},"page":"859","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Leveraging Immunofocusing and Virus-like Particle Display to Enhance Antibody Responses to the Malaria Blood-Stage Invasion Complex Antigen PfCyRPA"],"prefix":"10.3390","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1339-8620","authenticated-orcid":false,"given":"Kasper H.","family":"Bj\u00f6rnsson","sequence":"first","affiliation":[{"name":"Centre for translational Medicine and Parasitology, Department of Immunology and Microbiology (ISIM), Faculty of Health and Medical Sciences, University of Copenhagen, 2200 Copenhagen, Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria R.","family":"Bassi","sequence":"additional","affiliation":[{"name":"Centre for translational Medicine and Parasitology, Department of Immunology and Microbiology (ISIM), Faculty of Health and Medical Sciences, University of Copenhagen, 2200 Copenhagen, Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anne S.","family":"Knudsen","sequence":"additional","affiliation":[{"name":"Centre for translational Medicine and Parasitology, Department of Immunology and Microbiology (ISIM), Faculty of Health and Medical Sciences, University of Copenhagen, 2200 Copenhagen, Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0068-7733","authenticated-orcid":false,"given":"Kara-Lee","family":"Aves","sequence":"additional","affiliation":[{"name":"Centre for translational Medicine and Parasitology, Department of Immunology and Microbiology (ISIM), Faculty of Health and Medical Sciences, University of Copenhagen, 2200 Copenhagen, Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"\u00c8lia","family":"Morella Roig","sequence":"additional","affiliation":[{"name":"Centre for translational Medicine and Parasitology, Department of Immunology and Microbiology (ISIM), Faculty of Health and Medical Sciences, University of Copenhagen, 2200 Copenhagen, Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8782-7830","authenticated-orcid":false,"given":"Adam F.","family":"Sander","sequence":"additional","affiliation":[{"name":"Centre for translational Medicine and Parasitology, Department of Immunology and Microbiology (ISIM), Faculty of Health and Medical Sciences, University of Copenhagen, 2200 Copenhagen, Denmark"},{"name":"AdaptVac, Ole Maal\u00f8es Vej 3, 2200 Copenhagen, Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6510-495X","authenticated-orcid":false,"given":"Lea","family":"Barfod","sequence":"additional","affiliation":[{"name":"Centre for translational Medicine and Parasitology, Department of Immunology and Microbiology (ISIM), Faculty of Health and Medical Sciences, University of Copenhagen, 2200 Copenhagen, Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,7,30]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2039","DOI":"10.1038\/s41564-022-01261-2","article-title":"PCRCR Complex Is Essential for Invasion of Human Erythrocytes by Plasmodium falciparum","volume":"7","author":"Scally","year":"2022","journal-title":"Nat. Microbiol."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Chen, L., Lopaticki, S., Riglar, D.T., Dekiwadia, C., Uboldi, A.D., Tham, W.H., O\u2019Neill, M.T., Richard, D., Baum, J., and Ralph, S.A. (2011). An EGF-like Protein Forms a Complex with PfRh5 and Is Required for Invasion of Human Erythrocytes by Plasmodium falciparum. PLoS Pathog., 7.","DOI":"10.1371\/journal.ppat.1002199"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"371","DOI":"10.1016\/j.ijpara.2008.10.006","article-title":"Reticulocyte-Binding Protein Homologue 5\u2014An Essential Adhesin Involved in Invasion of Human Erythrocytes by Plasmodium falciparum","volume":"39","author":"Baum","year":"2009","journal-title":"Int. J. Parasitol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"6225","DOI":"10.4049\/jimmunol.1103177","article-title":"Passive Immunoprotection of Plasmodium falciparum-Infected Mice Designates the CyRPA as Candidate Malaria Vaccine Antigen","volume":"188","author":"Dreyer","year":"2012","journal-title":"J. Immunol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1179","DOI":"10.1073\/pnas.1415466112","article-title":"Multiprotein Complex between the GPI-Anchored CyRPA with PfRH5 and PfRipr Is Crucial for Plasmodium falciparum Erythrocyte Invasion","volume":"112","author":"Reddy","year":"2015","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"60","DOI":"10.1016\/j.chom.2016.06.004","article-title":"Essential Role of the PfRh5\/PfRipr\/CyRPA Complex during Plasmodium falciparum Invasion of Erythrocytes","volume":"20","author":"Volz","year":"2016","journal-title":"Cell Host Microbe"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"118","DOI":"10.1038\/s41586-018-0779-6","article-title":"Structure of Plasmodium falciparum Rh5\u2013CyRPA\u2013Ripr Invasion Complex","volume":"565","author":"Wong","year":"2019","journal-title":"Nature"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"534","DOI":"10.1038\/nature10606","article-title":"Basigin Is a Receptor Essential for Erythrocyte Invasion by Plasmodium falciparum","volume":"480","author":"Crosnier","year":"2011","journal-title":"Nature"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Waweru, H., Kanoi, B.N., Kuja, J.O., Maranga, M., Kongere, J., Maina, M., Kinyua, J., and Gitaka, J. (2023). Limited Genetic Variations of the Rh5-CyRPA-Ripr Invasion Complex in Plasmodium falciparum Parasite Population in Selected Malaria-Endemic Regions, Kenya. Front. Trop. Dis., 4.","DOI":"10.3389\/fitd.2023.1102265"},{"key":"ref_10","unstructured":"MalariaGEN, Ahouidi, A., Ali, M., Almagro-Garcia, J., Amambua-Ngwa, A., Amaratunga, C., Amato, R., Amenga-Etego, L., Andagalu, B., and Anderson, T.J.C. (2021). An Open Dataset of Plasmodium falciparum Genome Variation in 7000 Worldwide Samples [version 2; peer review: 2 approved]. Wellcome Open Res., 6, 42."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"545","DOI":"10.1016\/j.pt.2020.04.003","article-title":"The RH5-CyRPA-Ripr Complex as a Malaria Vaccine Target","volume":"36","author":"Ragotte","year":"2020","journal-title":"Trends Parasitol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"373","DOI":"10.1016\/j.vaccine.2012.10.106","article-title":"A Full-Length Recombinant Plasmodium falciparum PfRH5 Protein Induces Inhibitory Antibodies That Are Effective across Common PfRH5 Genetic Variants","volume":"31","author":"Bustamante","year":"2013","journal-title":"Vaccine"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"152","DOI":"10.1128\/IAI.00970-13","article-title":"Bacterially Expressed Full-Length Recombinant Plasmodium falciparum RH5 Protein Binds Erythrocytes and Elicits Potent Strain-Transcending Parasite-Neutralizing Antibodies","volume":"82","author":"Pandey","year":"2014","journal-title":"Infect. Immun."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Healer, J., Wong, W., Thompson, J.K., He, W., Birkinshaw, R.W., Miura, K., Long, C.A., Soroka, V., S\u00f8gaard, T.M.M., and J\u00f8rgensen, T. (2019). Neutralising Antibodies Block the Function of Rh5\/Ripr\/CyRPA Complex during Invasion of Plasmodium falciparum into Human Erythrocytes. Cell Microbiol., 21.","DOI":"10.1111\/cmi.13030"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"5612","DOI":"10.1016\/j.vaccine.2016.09.028","article-title":"Identification of Plasmodium falciparum Reticulocyte Binding Protein Homologue 5-Interacting Protein, PfRipr, as a Highly Conserved Blood-Stage Malaria Vaccine Candidate","volume":"34","author":"Ntege","year":"2016","journal-title":"Vaccine"},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Knudsen, A.S., Bj\u00f6rnsson, K.H., Bassi, M.R., Walker, M.R., Kok, A., Cristinoi, B., Jensen, A.R., and Barfod, L. (2021). Strain-Dependent Inhibition of Erythrocyte Invasion by Monoclonal Antibodies Against Plasmodium falciparum CyRPA. Front. Immunol., 12.","DOI":"10.3389\/fimmu.2021.716305"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"3040","DOI":"10.1038\/s41598-022-06921-1","article-title":"Enhancing Neutralization of Plasmodium falciparum Using a Novel Monoclonal Antibody against the Rhoptry-Associated Membrane Antigen","volume":"12","author":"Knudsen","year":"2022","journal-title":"Sci. Rep."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"e21347","DOI":"10.7554\/eLife.21347","article-title":"Structural Basis for Inhibition of Erythrocyte Invasion by Antibodies to Plasmodium falciparum Protein CyRPA","volume":"6","author":"Chen","year":"2017","journal-title":"Elife"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"e20383","DOI":"10.7554\/eLife.20383","article-title":"Structure of the Malaria Vaccine Candidate Antigen CyRPA and Its Complex with a Parasite Invasion Inhibitory Antibody","volume":"6","author":"Favuzza","year":"2017","journal-title":"Elife"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"245","DOI":"10.4049\/jimmunol.1302045","article-title":"Neutralization of Plasmodium falciparum Merozoites by Antibodies against PfRH5","volume":"192","author":"Douglas","year":"2014","journal-title":"J. Immunol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"601","DOI":"10.1038\/ncomms1615","article-title":"The Blood-Stage Malaria Antigen PfRH5 Is Susceptible to Vaccine-Inducible Cross-Strain Neutralizing Antibody","volume":"2","author":"Douglas","year":"2011","journal-title":"Nat. Commun."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"e96381","DOI":"10.1172\/jci.insight.96381","article-title":"Human Vaccination against RH5 Induces Neutralizing Antimalarial Antibodies That Inhibit RH5 Invasion Complex Interactions","volume":"2","author":"Payne","year":"2017","journal-title":"JCI Insight"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Healer, J., Thompson, J.K., Mackwell, K.L., Browne, C.D., Seager, B.A., Ngo, A., Lowes, K.N., Silk, S.E., Pulido, D., and King, L.D.W. (2022). RH5.1-CyRPA-Ripr Antigen Combination Vaccine Shows Little Improvement over RH5.1 in a Preclinical Setting. Front. Cell Infect. Microbiol., 12.","DOI":"10.3389\/fcimb.2022.1049065"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"130","DOI":"10.1016\/j.chom.2014.11.017","article-title":"A PfRH5-Based Vaccine Is Efficacious against Heterologous Strain Blood-Stage Plasmodium falciparum Infection in Aotus Monkeys","volume":"17","author":"Douglas","year":"2015","journal-title":"Cell Host Microbe"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1953","DOI":"10.1038\/s41467-019-09894-4","article-title":"A Defined Mechanistic Correlate of Protection against Plasmodium falciparum Malaria in Non-Human Primates","volume":"10","author":"Douglas","year":"2019","journal-title":"Nat. Commun."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"701","DOI":"10.1016\/j.medj.2021.03.014","article-title":"Reduced Blood-Stage Malaria Growth and Immune Correlates in Humans Following RH5 Vaccination","volume":"2","author":"Minassian","year":"2021","journal-title":"Med"},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Fernandes, B., Sousa, M., Castro, R., Sch\u00e4fer, A., Hauser, J., Schulze, K., Amacker, M., Tamborrini, M., Pluschke, G., and Alves, P.M. (2022). Scalable Process for High-Yield Production of PfCyRPA Using Insect Cells for Inclusion in a Malaria Virosome-Based Vaccine Candidate. Front. Bioeng. Biotechnol., 10.","DOI":"10.3389\/fbioe.2022.879078"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"933","DOI":"10.1038\/s41467-022-28601-4","article-title":"Heterotypic Interactions Drive Antibody Synergy against a Malaria Vaccine Candidate","volume":"13","author":"Ragotte","year":"2022","journal-title":"Nat. Commun."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"3677","DOI":"10.1128\/IAI.73.6.3677-3685.2005","article-title":"Phase 1 Clinical Trial of Apical Membrane Antigen 1: An Asexual Blood-Stage Vaccine for Plasmodium falciparum Malaria","volume":"73","author":"Malkin","year":"2005","journal-title":"Infect. Immun."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"11802","DOI":"10.1038\/s41598-020-67877-8","article-title":"Bliss\u2019 and Loewe\u2019s Additive and Synergistic Effects in Plasmodium falciparum Growth Inhibition by AMA1-RON2L, RH5, RIPR and CyRPA Antibody Combinations","volume":"10","author":"Azasi","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"787","DOI":"10.1038\/nri2868","article-title":"Vaccine Delivery: A Matter of Size, Geometry, Kinetics and Molecular Patterns","volume":"10","author":"Bachmann","year":"2010","journal-title":"Nat. Rev. Immunol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"3305","DOI":"10.1002\/1521-4141(200211)32:11<3305::AID-IMMU3305>3.0.CO;2-J","article-title":"Regulation of IgG Antibody Responses by Epitope Density and CD21-Mediated Costimulation","volume":"32","author":"Jegerlehner","year":"2002","journal-title":"Eur. J. Immunol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1448","DOI":"10.1126\/science.8248784","article-title":"The Influence of Antigen Organization on B Cell Responsiveness","volume":"262","author":"Bachmann","year":"1993","journal-title":"Science"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"4768","DOI":"10.1016\/j.vaccine.2017.12.079","article-title":"Explanations for the High Potency of HPV Prophylactic Vaccines","volume":"36","author":"Schiller","year":"2018","journal-title":"Vaccine"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"E690","DOI":"10.1073\/pnas.1115485109","article-title":"Peptide Tag Forming a Rapid Covalent Bond to a Protein, through Engineering a Bacterial Adhesin","volume":"109","author":"Zakeri","year":"2012","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Thrane, S., Janitzek, C.M., Matondo, S., Resende, M., Gustavsson, T., Jongh, W.A., Clemmensen, S., Roeffen, W., Vegte-Bolmer, M., and Gemert, G.J. (2016). Bacterial Superglue Enables Easy Development of Efficient Virus-like Particle Based Vaccines. J. Nanobiotechnol., 14.","DOI":"10.1186\/s12951-016-0181-1"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"324","DOI":"10.1038\/s41467-020-20251-8","article-title":"Capsid-like Particles Decorated with the SARS-CoV-2 Receptor-Binding Domain Elicit Strong Virus Neutralization Activity","volume":"12","author":"Fougeroux","year":"2021","journal-title":"Nat. Commun."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"8855","DOI":"10.1021\/acsnano.8b02805","article-title":"Engineering a Rugged Nanoscaffold to Enhance Plug-and-Display Vaccination","volume":"12","author":"Bruun","year":"2018","journal-title":"ACS Nano"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"e00245-23","DOI":"10.1128\/iai.00245-23","article-title":"Development of a Tag\/Catcher-Mediated Capsid Virus-like Particle Vaccine Presenting the Conserved Neisseria Gonorrhoeae SliC Antigen That Blocks Human Lysozyme","volume":"91","author":"Martinez","year":"2023","journal-title":"Infect. Immun."},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Goks\u00f8yr, L., Skrzypczak, M., Sampson, M., Nielsen, M.A., Salanti, A., Theander, T.G., Remaley, A.T., De Jongh, W.A., and Sander, A.F. (2023). A CVLP-Based Vaccine Displaying Full-Length PCSK9 Elicits a Higher Reduction in Plasma PCSK9 Than Similar Peptide-Based CVLP Vaccines. Vaccines, 11.","DOI":"10.3390\/vaccines11010002"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"e140","DOI":"10.1016\/S2666-5247(22)00337-8","article-title":"First-in-Human Use of a Modular Capsid Virus-like Vaccine Platform: An Open-Label, Non-Randomised, Phase 1 Clinical Trial of the SARS-CoV-2 Vaccine ABNCoV2","volume":"4","author":"Smit","year":"2023","journal-title":"Lancet Microbe"},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Aves, K.L., Janitzek, C.M., Fougeroux, C.E., Theander, T.G., and Sander, A.F. (2022). Freeze-Drying of a Capsid Virus-like Particle-Based Platform Allows Stable Storage of Vaccines at Ambient Temperature. Pharmaceutics, 14.","DOI":"10.3390\/pharmaceutics14061301"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"469","DOI":"10.1084\/jem.20151960","article-title":"Reverse Vaccinology 2.0: Human Immunology Instructs Vaccine Antigen Design","volume":"213","author":"Rappuoli","year":"2016","journal-title":"J. Exp. Med."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"191","DOI":"10.1016\/0022-1759(90)90029-U","article-title":"Removal of Endotoxin from Protein Solutions by Phase Separation Using Triton X-114","volume":"132","author":"Aida","year":"1990","journal-title":"J. Oflmmunol. Methods"},{"key":"ref_45","unstructured":"Iglewicz, B., and Hoaglin, D.C. (1993). How to Detect and Handle Outliers, ASQC Quality Press."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"4621","DOI":"10.1093\/nar\/21.19.4621","article-title":"Encapsidation of Heterologous RNAs by Bacteriophage MS2 Coat Protein","volume":"21","author":"Pickett","year":"1993","journal-title":"Nucleic Acids Res."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"4267","DOI":"10.1016\/j.jmb.2016.08.025","article-title":"Structure of AP205 Coat Protein Reveals Circular Permutation in SsRNA Bacteriophages","volume":"428","author":"Shishovs","year":"2016","journal-title":"J. Mol. Biol."},{"key":"ref_48","doi-asserted-by":"crossref","unstructured":"Chang, X., Krenger, P., Krueger, C.C., Zha, L., Han, J., Yermanos, A., Roongta, S., Mohsen, M.O., Oxenius, A., and Vogel, M. (2022). TLR7 Signaling Shapes and Maintains Antibody Diversity Upon Virus-Like Particle Immunization. Front. Immunol., 12.","DOI":"10.3389\/fimmu.2021.827256"},{"key":"ref_49","doi-asserted-by":"crossref","unstructured":"Thrane, S., Aves, K.L., Uddb\u00e4ck, I.E.M., Janitzek, C.M., Han, J., Yang, Y.R., Ward, A.B., Theander, T.G., Nielsen, M.A., and Salanti, A. (2020). A Vaccine Displaying a Trimeric Influenza-A HA Stem Protein on Capsid-Like Particles Elicits Potent and Long-Lasting Protection in Mice. Vaccines, 8.","DOI":"10.3390\/vaccines8030389"},{"key":"ref_50","doi-asserted-by":"crossref","unstructured":"Harrison, T.E., Alam, N., Farrell, B., Quinkert, D., Lias, A.M., King, L.D.W., Draper, S.J., Campeotto, I., and Higgins, M.K. (2024). Rational Structure-Guided Design of a Blood Stage Malaria Vaccine Immunogen Presenting a Single Epitope from PfRH5. bioRxiv.","DOI":"10.1101\/2024.02.29.582763"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"4857","DOI":"10.1038\/s41467-024-48721-3","article-title":"Development of an Improved Blood-Stage Malaria Vaccine Targeting the Essential RH5-CyRPA-RIPR Invasion Complex","volume":"15","author":"Williams","year":"2024","journal-title":"Nat. Commun."}],"container-title":["Vaccines"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-393X\/12\/8\/859\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T15:26:44Z","timestamp":1760110004000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-393X\/12\/8\/859"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,7,30]]},"references-count":51,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2024,8]]}},"alternative-id":["vaccines12080859"],"URL":"https:\/\/doi.org\/10.3390\/vaccines12080859","relation":{},"ISSN":["2076-393X"],"issn-type":[{"value":"2076-393X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,7,30]]}}}