{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,19]],"date-time":"2026-09-19T17:41:04Z","timestamp":1789839664113,"version":"4.0.1"},"reference-count":23,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2020,6,9]],"date-time":"2020-06-09T00:00:00Z","timestamp":1591660800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2020,6,9]],"date-time":"2020-06-09T00:00:00Z","timestamp":1591660800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/501100012246","name":"Priority Academic Program Development of Jiangsu Higher Education Institutions","doi-asserted-by":"publisher","award":["PAPD"],"award-info":[{"award-number":["PAPD"]}],"id":[{"id":"10.13039\/501100012246","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100012165","name":"Key Technologies Research and Development Program","doi-asserted-by":"publisher","award":["2018YFC0840404-3"],"award-info":[{"award-number":["2018YFC0840404-3"]}],"id":[{"id":"10.13039\/501100012165","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Virol J"],"published-print":{"date-parts":[[2020,12]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec>\n<jats:title>Background<\/jats:title>\n<jats:p>Porcine circovirus type 2 (PCV2) is an economically important pathogen affecting swine industry worldwide. The production of current PCV2 vaccines is time-consuming and expensive. Elastin-like polypeptides (ELP) undergo temperature-dependent inverse phase transition and ELPylated proteins can be purified simply by inverse transition cycling (ITC).<\/jats:p>\n<\/jats:sec><jats:sec>\n<jats:title>Methods<\/jats:title>\n<jats:p>The Cap protein of PCV2b, together with the virus neutralizing (VN) epitopes of PCV2a, PCV2d and PCV2e, was expressed in <jats:italic>E. coli<\/jats:italic> as an ELPylated protein, and purified by ITC in the presence of mild detergents. For the control purpose, the Cap protein was also expressed as a His-tagged protein and purified by nickel affinity chromatography. The formation of ELPylated VLP (ELP-VLP) and His-tagged VLP (VLP) was revealed by transmission electron microscopy. Mice were immunized two times with the two forms of VLP and the antigen-specific IgG antibody, VN antibody, cytokine responses and immunoprotection against PCV2 challenge were compared.<\/jats:p>\n<\/jats:sec><jats:sec>\n<jats:title>Results<\/jats:title>\n<jats:p>ELPylated Cap protein was expressed as a soluble protein and purified to 94.3% purity by ITC in the presence of 1% Triton X-100 and 0.5\u2009M urea. His-tagged Cap fusion protein was expressed as insoluble inclusion bodies and purified to 90% purity under denatured conditions. The two purified fusion proteins assembled into VLP with similar morphology. Compared to immunization with VLP, immunization with ELP-VLP induced significantly (<jats:italic>p<\/jats:italic>\u2009&lt;\u20090.01) stronger VN antibody response and slightly (<jats:italic>p<\/jats:italic>\u2009&lt;\u20090.05) stronger Cap-specific IgG antibody response, cytokine production and immunoprotection against PCV2 challenge.<\/jats:p>\n<\/jats:sec><jats:sec>\n<jats:title>Conclusion<\/jats:title>\n<jats:p>A novel ELPylation platform for easy preparation of PCV2 VLP was established and the prepared ELP-VLP was more immunogenic than VLP. The ELPylation technology could be used for other VLP preparation and the prepared ELP-VLP could be developed as a novel PCV2 subunit vaccine.<\/jats:p>\n<\/jats:sec>","DOI":"10.1186\/s12985-020-01346-6","type":"journal-article","created":{"date-parts":[[2020,6,9]],"date-time":"2020-06-09T13:20:04Z","timestamp":1591708804000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":12,"title":["Generation and immunogenicity assessment of ELPylated virus-like particles of porcine circovirus type 2"],"prefix":"10.1186","volume":"17","author":[{"given":"Yangyang","family":"Li","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yajie","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jian","family":"Cheng","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaohui","family":"Zhou","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huipeng","family":"Lu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinyu","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaoli","family":"Xia","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huaichang","family":"Sun","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2020,6,9]]},"reference":[{"key":"1346_CR1","doi-asserted-by":"publisher","first-page":"326","DOI":"10.1016\/j.tvjl.2004.01.012","volume":"169","author":"C Chae","year":"2005","unstructured":"Chae C. A review of porcine circovirus 2-associated syndromes and diseases. Vet J. 2005;169:326\u201336.","journal-title":"Vet J"},{"key":"1346_CR2","doi-asserted-by":"publisher","first-page":"591","DOI":"10.1177\/104063870701900601","volume":"19","author":"T Opriessnig","year":"2007","unstructured":"Opriessnig T, Meng XJ, Halbur PG. Porcine circovirus type 2 associated disease: update on current terminology, clinical manifestations, pathogenesis, diagnosis, and intervention strategies. J Vet Diagn Investig. 2007;19:591\u2013615.","journal-title":"J Vet Diagn Investig"},{"key":"1346_CR3","doi-asserted-by":"publisher","first-page":"151","DOI":"10.1016\/j.tvjl.2012.06.031","volume":"194","author":"C Chae","year":"2012","unstructured":"Chae C. Commercial porcine circovirus type 2 vaccines: efficacy and clinical application. Vet J. 2012;194:151\u20137.","journal-title":"Vet J"},{"key":"1346_CR4","doi-asserted-by":"publisher","first-page":"423","DOI":"10.1016\/j.virol.2007.08.014","volume":"369","author":"Y Zhu","year":"2007","unstructured":"Zhu Y, Lau A, Lau J, Jia Q, Karuppannan AK, Kwang J. Enhanced replication of porcine circovirus type 2 (PCV2) in a homogeneous subpopulation of PK15 cell line. Virol. 2007;369:423\u201330.","journal-title":"Virol."},{"issue":"Suppl","key":"1346_CR5","doi-asserted-by":"publisher","first-page":"S42","DOI":"10.1016\/j.jip.2011.05.004","volume":"107","author":"T Vicente","year":"2011","unstructured":"Vicente T, Rold\u00e3o A, Peixoto C, Carrondo MJ, Alves PM. Large-scale production and purification of VLP-based vaccines. J Invertebr Pathol. 2011;107(Suppl):S42\u20138.","journal-title":"J Invertebr Pathol"},{"key":"1346_CR6","doi-asserted-by":"publisher","first-page":"78","DOI":"10.1016\/j.jbiotec.2016.01.017","volume":"220","author":"PC Wu","year":"2016","unstructured":"Wu PC, Chen TY, Chi JN, Chien MS, Huang C. Efficient expression and purification of porcine circovirus type 2 virus-like particles in Escherichia coli. J Biotechnol. 2016;220:78\u201385.","journal-title":"J Biotechnol"},{"key":"1346_CR7","doi-asserted-by":"publisher","first-page":"3650","DOI":"10.1002\/art.22952","volume":"56","author":"MF Shamji","year":"2007","unstructured":"Shamji MF, Betre H, Kraus VB, Chen J, Chilkoti A, Pichika R, Masuda K, Setton LA. Development and characterization of a fusion protein between thermally responsive elastin-like polypeptide and interleukin-1 receptor antagonist. Arthritis Rheum. 2007;56:3650\u201361.","journal-title":"Arthritis Rheum"},{"key":"1346_CR8","doi-asserted-by":"publisher","first-page":"23","DOI":"10.1016\/0079-6107(92)90003-O","volume":"57","author":"DW Urry","year":"1992","unstructured":"Urry DW. Free energy transduction in polypeptides and proteins based on inverse temperature transitions. Prog Biophys Mol Biol. 1992;57:23\u201357.","journal-title":"Prog Biophys Mol Biol"},{"key":"1346_CR9","doi-asserted-by":"publisher","first-page":"1112","DOI":"10.1038\/15100","volume":"17","author":"DE Meyer","year":"1999","unstructured":"Meyer DE, Chilkoti A. Purification of recombinant proteins by fusion with thermally-responsive polypeptides. Nat Biotechnol. 1999;17:1112\u20135.","journal-title":"Nat Biotechnol"},{"key":"1346_CR10","doi-asserted-by":"publisher","first-page":"1456","DOI":"10.1016\/j.addr.2010.05.004","volume":"62","author":"JR McDaniel","year":"2010","unstructured":"McDaniel JR, Callahan DJ, Chilkoti A. Drug delivery to solid tumors by elastin-like polypeptides. Adv Drug Deliv Rev. 2010;62:1456\u201367.","journal-title":"Adv Drug Deliv Rev"},{"key":"1346_CR11","doi-asserted-by":"publisher","first-page":"101","DOI":"10.1111\/jnc.12782","volume":"131","author":"SM Hearst","year":"2014","unstructured":"Hearst SM, Shao Q, Lopez M, Raucher D, Vig PJS. The design and delivery of PKA inhibitory polypeptide to treat SCA1. J Neurochem. 2014;131:101\u20134.","journal-title":"J Neurochem"},{"key":"1346_CR12","doi-asserted-by":"publisher","first-page":"1617","DOI":"10.1002\/bit.25998","volume":"113","author":"A Yeboah","year":"2016","unstructured":"Yeboah A, Cohen RI, Rabolli C, Yarmush ML, Berthiaume F. Elastin-like polypeptides: a strategic fusion partner for biologics. Biotechnol Bioeng. 2016;113:1617\u201327.","journal-title":"Biotechnol Bioeng"},{"key":"1346_CR13","doi-asserted-by":"publisher","first-page":"1450004","DOI":"10.1142\/S1793984414500044","volume":"4","author":"RS Ingrole","year":"2014","unstructured":"Ingrole RS, Tao W, Tripathy JN, Gill HS. Synthesis and immunogenicity assessment of elastin-like polypeptide-M2e construct as an influenza antigen. Nano Life. 2014;4:1450004.","journal-title":"Nano Life"},{"key":"1346_CR14","doi-asserted-by":"publisher","first-page":"582","DOI":"10.1111\/pbi.12049","volume":"11","author":"HT Phan","year":"2013","unstructured":"Phan HT, Pohl J, Floss DM, Rabenstein F, Veits J, Le BT, Chu HH, Hause G, Mettenleiter T, Conrad U. ELPylated haemagglutinin produced in tobacco plants induce potentially neutralizing antibodies against H5N1 viruses in mice. Plant Biotechnol J. 2013;11:582\u201393.","journal-title":"Plant Biotechnol J"},{"key":"1346_CR15","doi-asserted-by":"publisher","first-page":"53","DOI":"10.1016\/j.pep.2018.08.012","volume":"153","author":"Y Zong","year":"2019","unstructured":"Zong Y, Tan X, Xiao J, Zhang X, Xia X, Sun H. Half-life extension of porcine interferon-\u03b1 by fusion to the IgG-binding domain of streptococcal G protein. Protein Expr Purif. 2019;153:53\u20138.","journal-title":"Protein Expr Purif"},{"key":"1346_CR16","doi-asserted-by":"publisher","first-page":"188","DOI":"10.1186\/1471-2180-11-188","volume":"11","author":"LP Huang","year":"2011","unstructured":"Huang LP, Lu YH, Wei YW, Guo LJ, Liu CM. Identification of one critical amino acid that determines a conformational neutralizing epitope in the capsid protein of porcine circovirus type 2. BMC Microbiol. 2011;11:188.","journal-title":"BMC Microbiol"},{"issue":"Web Server issu","key":"1346_CR17","doi-asserted-by":"publisher","first-page":"W526","DOI":"10.1093\/nar\/gki376","volume":"33","author":"A Grote","year":"2005","unstructured":"Grote A, Hiller K, Scheer M, M\u00fcnch R, N\u00f6rtemann B, Hempel DC, Jahn D. JCat: a novel tool to adapt codon usage of a target gene to its potential expression host. Nucleic Acids Res. 2005;33(Web Server issue):W526\u201331.","journal-title":"Nucleic Acids Res"},{"key":"1346_CR18","doi-asserted-by":"publisher","first-page":"18","DOI":"10.1016\/j.pep.2014.02.016","volume":"98","author":"WJ Liu","year":"2014","unstructured":"Liu WJ, Wu Q, Xu B, Zhang XY, Xia XL, Sun HC. Single-step purification of recombinant proteins using elastin-like peptide-mediated inverse transition cycling and self-processing module from Neisseria meningitides FrpC. Protein Expr Purif. 2014;98:18\u201324.","journal-title":"Protein Expr Purif"},{"key":"1346_CR19","doi-asserted-by":"publisher","first-page":"86","DOI":"10.1016\/j.pep.2016.08.013","volume":"128","author":"GY Li","year":"2016","unstructured":"Li GY, Xiao ZZ, Lu HP, Li YY, Zhou XH, Tan X, Zhang XY, Xia XL, Sun HC. A simple method for recombinant protein purification using self-aggregating peptide-tagged tobacco etch virus protease. Protein Expr Purif. 2016;128:86\u201392.","journal-title":"Protein Expr Purif"},{"issue":"2","key":"1346_CR20","doi-asserted-by":"publisher","first-page":"143","DOI":"10.1089\/vim.2000.13.143","volume":"13","author":"RM Pogranichnyy","year":"2000","unstructured":"Pogranichnyy RM, Yoon KJ, Harms PA, Swenson SL, Zimmerman JJ, Sorden SD. Characterization of immune response of young pigs to porcine circovirus type 2 infection. Viral Immunol. 2000;13(2):143\u201353.","journal-title":"Viral Immunol"},{"key":"1346_CR21","doi-asserted-by":"publisher","first-page":"55","DOI":"10.1002\/bip.22137","volume":"99","author":"A Thapa","year":"2013","unstructured":"Thapa A, Han W, Simons RH, Chilkoti A, Chi EY, L\u00f3pez GP. Effect of detergents on the thermal behavior of elastin-like polypeptides. Biopolymers. 2013;99:55\u201362.","journal-title":"Biopolymers."},{"key":"1346_CR22","doi-asserted-by":"publisher","first-page":"379","DOI":"10.1111\/j.1467-7652.2008.00326.x","volume":"6","author":"DM Floss","year":"2008","unstructured":"Floss DM, Sack M, Stadlmann J, Rademacher T, Scheller J, Stoger E, Fischer R, Conrad U. Biochemical and functional characterization of anti-HIV antibody-ELP fusion proteins from transgenic plants. Plant Biotechnol J. 2008;6:379.","journal-title":"Plant Biotechnol J"},{"key":"1346_CR23","doi-asserted-by":"publisher","first-page":"23","DOI":"10.1002\/ame2.12009","volume":"1","author":"T Ouyang","year":"2018","unstructured":"Ouyang T, Liu XH, Ouyang HS, Ren LZ. Mouse models of porcine circovirus 2 infection. Animal Model Exp Med. 2018;1:23\u20138.","journal-title":"Animal Model Exp Med"}],"container-title":["Virology Journal"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s12985-020-01346-6.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/article\/10.1186\/s12985-020-01346-6\/fulltext.html","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s12985-020-01346-6.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,6,8]],"date-time":"2021-06-08T23:33:11Z","timestamp":1623195191000},"score":1,"resource":{"primary":{"URL":"https:\/\/virologyj.biomedcentral.com\/articles\/10.1186\/s12985-020-01346-6"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,6,9]]},"references-count":23,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2020,12]]}},"alternative-id":["1346"],"URL":"https:\/\/doi.org\/10.1186\/s12985-020-01346-6","relation":{},"ISSN":["1743-422X"],"issn-type":[{"value":"1743-422X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,6,9]]},"assertion":[{"value":"25 August 2019","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"2 June 2020","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"9 June 2020","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"This research, including the procedure and protocol of animal immunization, sample collection and processing, was carried out according to the recommendations in the Guide for the Care and Use of Laboratory Animals of the Yangzhou University. The protocol was approved by Medical Experimental Animal Center of Jiangsu Province (Permit Number: 2140880).","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval and consent to participate"}},{"value":"Not applicable.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Consent for publication"}},{"value":"The authors declare that they have no conflict of interest.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"72"}}