{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T19:15:25Z","timestamp":1792782925548,"version":"4.5.0"},"reference-count":40,"publisher":"Springer Science and Business Media LLC","issue":"5","license":[{"start":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T00:00:00Z","timestamp":1789689600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T00:00:00Z","timestamp":1789689600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/501100002322","name":"Coordena\u00e7\u00e3o de Aperfei\u00e7oamento de Pessoal de N\u00edvel Superior","doi-asserted-by":"publisher","award":["001"],"award-info":[{"award-number":["001"]}],"id":[{"id":"10.13039\/501100002322","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004263","name":"Funda\u00e7\u00e3o de Amparo \u00e0 Pesquisa do Estado do Rio Grande do Sul","doi-asserted-by":"publisher","award":["Edital 07\/2021"],"award-info":[{"award-number":["Edital 07\/2021"]}],"id":[{"id":"10.13039\/501100004263","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Universidade Federal De Pelotas","id":[{"id":"https:\/\/ror.org\/05msy9z54","id-type":"ROR","asserted-by":"crossref"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Biotechnol Lett"],"published-print":{"date-parts":[[2026,10]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Proliferative enteropathy is a gastrointestinal disease caused by the intracellular bacterium\n                    <jats:italic>Lawsonia intracellularis<\/jats:italic>\n                    , affecting swine populations worldwide and leading to significant economic losses due to reduced growth performance, increased mortality, and treatment expenses. Current vaccines present limitations related to production processes, stability, and overall cost. This study aimed to design and evaluat chimeric proteins containing immunogenic epitopes of\n                    <jats:italic>L. intracellularis<\/jats:italic>\n                    . A recombinant chimera, designated Li_full, was rationally designed in silico by integrating predicted B-cell and MHC epitopes relevant to swine immunity. To further investigate the immunogenic potential of distinct regions, Li_full was subdivided into three fractions (Li_1, Li_2, and Li_3). All constructs were cloned into an\n                    <jats:italic>Escherichia coli<\/jats:italic>\n                    expression system, allowing efficient production and biochemical characterization of the recombinant proteins. Antigenicity was confirmed by strong reactivity with sera from naturally infected pigs. Immunogenicity was assessed through vaccination in mice, which developed significantly (p &lt; 0.05) elevated antibody titers, demonstrating the ability of the chimeras to elicit robust humoral immune responses. The integration of bioinformatics-driven epitope prediction with recombinant DNA technology represents a promising and innovative strategy for controlling proliferative enteropathy. This study underscores the value of in silico design strategies for the development of targeted veterinary immunogens and provides a foundation for further research aimed at evaluating protective efficacy under field conditions.\n                  <\/jats:p>","DOI":"10.1007\/s10529-026-03786-6","type":"journal-article","created":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T06:20:35Z","timestamp":1789712435000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Design and functional characterization of recombinant chimeras as vaccine candidates against Lawsonia intracellularis"],"prefix":"10.1007","volume":"48","author":[{"given":"Ana Vit\u00f3ria","family":"Costa","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Neida L.","family":"Conrad","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jo\u00e3o Pedro G.","family":"Greco","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Frederico S.","family":"Kremer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0941-7286","authenticated-orcid":false,"given":"F\u00e1bio P. L.","family":"Leite","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,9,18]]},"reference":[{"key":"3786_CR1","doi-asserted-by":"publisher","first-page":"207","DOI":"10.3390\/vaccines13020207","volume":"13","author":"KL Aves","year":"2025","unstructured":"Aves KL, Fresno AH, Nisar S, Saraiva MM, Goecke NB, Sander AF, Nielsen MA, Olsen JE, Guerra PR (2025) Outer membrane proteins as vaccine targets against Lawsonia intracellularis in piglets. Vaccines (Basel) 13:207. https:\/\/doi.org\/10.3390\/vaccines13020207","journal-title":"Vaccines (Basel)"},{"key":"3786_CR2","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetvac.2025.100132","volume":"4","author":"NL Conrad","year":"2025","unstructured":"Conrad NL, Gon\u00e7alves Zorzi VS, Mazzoleni I, Costa AV, Eug\u00eanio Ara\u00fajo Piraine R, Leite FPL (2025) Immunogenicity of a recombinant vaccine against Lawsonia intracellularis in mice. Veterinary Vaccine 4:100132. https:\/\/doi.org\/10.1016\/j.vetvac.2025.100132","journal-title":"Veterinary Vaccine"},{"key":"3786_CR3","doi-asserted-by":"publisher","DOI":"10.1016\/j.rvsc.2026.106132","volume":"204","author":"NL Conrad","year":"2026","unstructured":"Conrad NL, Mazzoleni I, Abreu MC, Costa AV, Di Gi\u00e1como CP, Zorzi VSG, Leite FPL (2026) Horse immune response of recombinant Lawsonia intracellularis vaccine: assessing the immunomodulatory impact of probiotic supplementation. Res Vet Sci 204:106132. https:\/\/doi.org\/10.1016\/j.rvsc.2026.106132","journal-title":"Res Vet Sci"},{"key":"3786_CR4","doi-asserted-by":"publisher","first-page":"1446","DOI":"10.2460\/javma.1998.212.09.1446","volume":"212","author":"DM Cooper","year":"1998","unstructured":"Cooper DM, Gebhart CJ (1998) Comparative aspects of proliferative enteritis. J Am Vet Med Assoc 212:1446\u20131451. https:\/\/doi.org\/10.2460\/javma.1998.212.09.1446","journal-title":"J Am Vet Med Assoc"},{"key":"3786_CR5","doi-asserted-by":"publisher","DOI":"10.1038\/ncomms16073","volume":"8","author":"M Enamorado","year":"2017","unstructured":"Enamorado M, Iborra S, Priego E, Cueto FJ, Quintana JA, Mart\u00ednez-Cano S, Mej\u00edas-P\u00e9rez E, Esteban M, Melero I, Hidalgo A, Sancho D (2017) Enhanced anti-tumour immunity requires the interplay between resident and circulating memory CD8+ T cells. Nat Commun 8:16073. https:\/\/doi.org\/10.1038\/ncomms16073","journal-title":"Nat Commun"},{"key":"3786_CR6","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetimm.2021.110256","volume":"237","author":"KR Fourie","year":"2021","unstructured":"Fourie KR, Choudhary P, Ng SH, Obradovic M, Brownlie R, Anand SK, Wilson HL (2021) Evaluation of immunogenicity and protection mediated by Lawsonia intracellularis subunit vaccines. Vet Immunol Immunopathol 237:110256. https:\/\/doi.org\/10.1016\/j.vetimm.2021.110256","journal-title":"Vet Immunol Immunopathol"},{"key":"3786_CR7","doi-asserted-by":"publisher","first-page":"571","DOI":"10.1385\/1-59259-890-0:571","volume-title":"The proteomics protocols handbook","author":"E Gasteiger","year":"2005","unstructured":"Gasteiger E, Hoogland C, Gattiker A, Duvaud S, Wilkins MR, Appel RD, Bairoch A (2005) Protein identification and analysis tools on the ExPASy server. In: Walker JM (ed) The proteomics protocols handbook. Humana Press, Totowa, pp 571\u2013607. https:\/\/doi.org\/10.1385\/1-59259-890-0:571"},{"key":"3786_CR8","doi-asserted-by":"publisher","first-page":"620","DOI":"10.1177\/0300985817698206","volume":"54","author":"RMC Guedes","year":"2017","unstructured":"Guedes RMC, Machuca MA, Quiroga MA, Pereira CER, Resende TP, Gebhart CJ (2017) Lawsonia intracellularis in pigs: progression of lesions and involvement of apoptosis. Vet Pathol 54:620\u2013628. https:\/\/doi.org\/10.1177\/0300985817698206","journal-title":"Vet Pathol"},{"key":"3786_CR9","doi-asserted-by":"publisher","first-page":"171","DOI":"10.1146\/annurev-animal-020518-115014","volume":"8","author":"SE Hammer","year":"2020","unstructured":"Hammer SE, Ho C-S, Ando A, Rogel-Gaillard C, Charles M, Tector M, Tector AJ, Lunney JK (2020) Importance of the major histocompatibility complex (swine leukocyte antigen) in swine health and biomedical research. Annu Rev Anim Biosci 8:171\u2013198. https:\/\/doi.org\/10.1146\/annurev-animal-020518-115014","journal-title":"Annu Rev Anim Biosci"},{"key":"3786_CR10","doi-asserted-by":"publisher","DOI":"10.1155\/2010\/297505","author":"Y He","year":"2010","unstructured":"He Y, Xiang Z, Mobley HLT (2010) Vaxign: the first web-based vaccine design program for reverse vaccinology and applications for vaccine development. J Biomed Biotechnol. https:\/\/doi.org\/10.1155\/2010\/297505","journal-title":"J Biomed Biotechnol"},{"key":"3786_CR11","unstructured":"Holtkamp D (2020) Perdas econ\u00f4micas associadas \u00e0 ile\u00edte. O Presente Rural \u2013 Su\u00ednos\/Peixes. https:\/\/opresenterural.com.br\/perdas-economicas-associadas-a-ileite\/. Accessed 24 March 2026"},{"key":"3786_CR12","doi-asserted-by":"publisher","first-page":"33","DOI":"10.1111\/j.1751-0813.2008.00372.x","volume":"87","author":"P Holyoake","year":"2009","unstructured":"Holyoake P, Collins A, Donahoo M, Lising R, Emery D (2009) Identifying obstacles to reducing the use of antibiotics to control porcine proliferative enteropathy. Aust Vet J 87:33\u201334. https:\/\/doi.org\/10.1111\/j.1751-0813.2008.00372.x","journal-title":"Aust Vet J"},{"key":"3786_CR13","doi-asserted-by":"publisher","first-page":"423","DOI":"10.2353\/ajpath.2006.050312","volume":"168","author":"D Huster","year":"2006","unstructured":"Huster D, Finegold MJ, Morgan CT, Burkhead JL, Nixon R, Vanderwerf SM, Gilliam CT, Lutsenko S (2006) Consequences of copper accumulation in the livers of the Atp7b\u2212\/\u2212 (Wilson disease gene) knockout mice. Am J Pathol 168:423\u2013434. https:\/\/doi.org\/10.2353\/ajpath.2006.050312","journal-title":"Am J Pathol"},{"key":"3786_CR14","doi-asserted-by":"publisher","first-page":"W24","DOI":"10.1093\/nar\/gkx346","volume":"45","author":"MC Jespersen","year":"2017","unstructured":"Jespersen MC, Peters B, Nielsen M, Marcatili P (2017) BepiPred-2.0: improving sequence-based B-cell epitope prediction using conformational epitopes. Nucleic Acids Res 45:W24\u2013W29. https:\/\/doi.org\/10.1093\/nar\/gkx346","journal-title":"Nucleic Acids Res"},{"key":"3786_CR15","doi-asserted-by":"publisher","first-page":"214","DOI":"10.1111\/j.0105-2896.2006.00391.x","volume":"211","author":"CA Klebanoff","year":"2006","unstructured":"Klebanoff CA, Gattinoni L, Restifo NP (2006) CD8+ T\u2010cell memory in tumor immunology and immunotherapy. Immunol Rev 211:214\u2013224. https:\/\/doi.org\/10.1111\/j.0105-2896.2006.00391.x","journal-title":"Immunol Rev"},{"key":"3786_CR16","doi-asserted-by":"publisher","DOI":"10.3389\/fvets.2016.00120","author":"AB Kruse","year":"2017","unstructured":"Kruse AB, De Knegt LV, Nielsen LR, Alban L (2017) No clear effect of initiating vaccination against common endemic infections on the amounts of prescribed antimicrobials for Danish weaner and finishing pigs during 2007\u20132013. Front Vet Sci. https:\/\/doi.org\/10.3389\/fvets.2016.00120","journal-title":"Front Vet Sci"},{"key":"3786_CR17","doi-asserted-by":"publisher","first-page":"77","DOI":"10.1053\/jcpa.1999.0347","volume":"122","author":"GHK Lawson","year":"2000","unstructured":"Lawson GHK, Gebhart CJ (2000) Proliferative enteropathy. J Comp Pathol 122:77\u2013100. https:\/\/doi.org\/10.1053\/jcpa.1999.0347","journal-title":"J Comp Pathol"},{"key":"3786_CR18","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetmic.2021.109274","volume":"263","author":"M Li","year":"2021","unstructured":"Li M, Xiao N, Li J, Lin H, Fan H (2021) Evaluation of immune efficacy of Omp2 protein against Lawsonia intracellularis in mice. Vet Microbiol 263:109274. https:\/\/doi.org\/10.1016\/j.vetmic.2021.109274","journal-title":"Vet Microbiol"},{"key":"3786_CR19","doi-asserted-by":"publisher","first-page":"248","DOI":"10.3390\/vaccines13030248","volume":"13","author":"J Li","year":"2025","unstructured":"Li J, Ju Y, Jiang M, Li S, Yang X-Y (2025) Epitope-based vaccines: the next generation of promising vaccines against bacterial infection. Vaccines 13:248. https:\/\/doi.org\/10.3390\/vaccines13030248","journal-title":"Vaccines"},{"key":"3786_CR20","doi-asserted-by":"publisher","first-page":"255","DOI":"10.1016\/0034-5288(95)90013-6","volume":"59","author":"S McOrist","year":"1995","unstructured":"McOrist S, Jasni S, Mackie RA, Berschneider HM, Rowland AC, Lawson GHK (1995) Entry of the bacterium ileal symbiont intracellularis into cultured enterocytes and its subsequent release. Res Vet Sci 59:255\u2013260. https:\/\/doi.org\/10.1016\/0034-5288(95)90013-6","journal-title":"Res Vet Sci"},{"key":"3786_CR21","doi-asserted-by":"publisher","first-page":"419","DOI":"10.1038\/nrmicro1901","volume":"6","author":"D Medini","year":"2008","unstructured":"Medini D, Serruto D, Parkhill J, Relman DA, Donati C, Moxon R, Falkow S, Rappuoli R (2008) Microbiology in the post-genomic era. Nat Rev Microbiol 6:419\u2013430. https:\/\/doi.org\/10.1038\/nrmicro1901","journal-title":"Nat Rev Microbiol"},{"key":"3786_CR22","doi-asserted-by":"publisher","first-page":"1340","DOI":"10.1016\/j.vaccine.2019.01.029","volume":"37","author":"R Montesino","year":"2019","unstructured":"Montesino R, Guti\u00e9rrez N, Camacho F, Farn\u00f3s O, Andrades S, Gonz\u00e1lez A, Acosta J, Cortez-San Mart\u00edn M, S\u00e1nchez O, Ruiz A, Toledo JR (2019) Multi-antigenic recombinant subunit vaccine against Lawsonia intracellularis: the etiological agent of porcine proliferative enteropathy. Vaccine 37:1340\u20131349. https:\/\/doi.org\/10.1016\/j.vaccine.2019.01.029","journal-title":"Vaccine"},{"key":"3786_CR23","doi-asserted-by":"publisher","first-page":"1007","DOI":"10.1110\/ps.0239403","volume":"12","author":"M Nielsen","year":"2003","unstructured":"Nielsen M, Lundegaard C, Worning P, Lauem\u00f8ller SL, Lamberth K, Buus S, Brunak S, Lund O (2003) Reliable prediction of T-cell epitopes using neural networks with novel sequence representations. Protein Sci 12:1007\u20131017. https:\/\/doi.org\/10.1110\/ps.0239403","journal-title":"Protein Sci"},{"key":"3786_CR24","doi-asserted-by":"publisher","first-page":"131","DOI":"10.1016\/j.vetmic.2013.02.004","volume":"164","author":"MG Nogueira","year":"2013","unstructured":"Nogueira MG, Collins AM, Donahoo M, Emery D (2013) Immunological responses to vaccination following experimental Lawsonia intracellularis virulent challenge in pigs. Vet Microbiol 164:131\u2013138. https:\/\/doi.org\/10.1016\/j.vetmic.2013.02.004","journal-title":"Vet Microbiol"},{"key":"3786_CR25","doi-asserted-by":"publisher","first-page":"270","DOI":"10.1016\/j.vetmic.2019.07.014","volume":"235","author":"M Obradovic","year":"2019","unstructured":"Obradovic M, Pasternak JA, Hon Ng S, Allan B, Brownlie R, Wilson HL (2019) Immunoproteomic analysis of Lawsonia intracellularis identifies candidate neutralizing antibody targets for use in subunit vaccine development. Vet Microbiol 235:270\u2013279. https:\/\/doi.org\/10.1016\/j.vetmic.2019.07.014","journal-title":"Vet Microbiol"},{"key":"3786_CR26","doi-asserted-by":"publisher","DOI":"10.4142\/jvs.2019.20.e24","volume":"20","author":"S Park","year":"2019","unstructured":"Park S, Won G, Lee JH (2019) An attenuated Salmonella vaccine secreting Lawsonia intracellularis immunogenic antigens confers dual protection against porcine proliferative enteropathy and salmonellosis in a murine model. J Vet Sci 20:e24. https:\/\/doi.org\/10.4142\/jvs.2019.20.e24","journal-title":"J Vet Sci"},{"key":"3786_CR27","doi-asserted-by":"publisher","first-page":"1816","DOI":"10.1126\/science.287.5459.1816","volume":"287","author":"M Pizza","year":"2000","unstructured":"Pizza M, Scarlato V, Masignani V, Giuliani MM, Aric\u00f2 B, Comanducci M, Jennings GT, Baldi L, Bartolini E, Capecchi B, Galeotti CL, Luzzi E, Manetti R, Marchetti E, Mora M, Nuti S, Ratti G, Santini L, Savino S, Scarselli M, Storni E, Zuo P, Broeker M, Hundt E, Knapp B, Blair E, Mason T, Tettelin H, Hood DW, Jeffries AC, Saunders NJ, Granoff DM, Venter JC, Moxon ER, Grandi G, Rappuoli R (2000) Identification of vaccine candidates against serogroup B meningococcus by whole-genome sequencing. Science 287:1816\u20131820. https:\/\/doi.org\/10.1126\/science.287.5459.1816","journal-title":"Science"},{"key":"3786_CR28","doi-asserted-by":"publisher","DOI":"10.1186\/s12917-015-0618-z","volume":"11","author":"TP Resende","year":"2015","unstructured":"Resende TP, Pereira CER, Gabardo MDP, Haddad JPA, Lobato ZIP, Guedes RMC (2015) Serological profile, seroprevalence and risk factors related to Lawsonia intracellularis infection in swine herds from Minas Gerais State, Brazil. BMC Vet Res 11:306. https:\/\/doi.org\/10.1186\/s12917-015-0618-z","journal-title":"BMC Vet Res"},{"key":"3786_CR29","doi-asserted-by":"publisher","first-page":"156","DOI":"10.1016\/j.vaccine.2014.10.084","volume":"33","author":"U Riber","year":"2015","unstructured":"Riber U, Heegaard PMH, Cordes H, St\u00e5hl M, Jensen TK, Jungersen G (2015) Vaccination of pigs with attenuated Lawsonia intracellularis induced acute phase protein responses and primed cell-mediated immunity without reduction in bacterial shedding after challenge. Vaccine 33:156\u2013162. https:\/\/doi.org\/10.1016\/j.vaccine.2014.10.084","journal-title":"Vaccine"},{"key":"3786_CR30","doi-asserted-by":"publisher","first-page":"1500","DOI":"10.1016\/j.vaccine.2017.12.049","volume":"36","author":"F Roerink","year":"2018","unstructured":"Roerink F, Morgan CL, Knetter SM, Passat M-H, Archibald AL, Ait-Ali T, Strait EL (2018) A novel inactivated vaccine against Lawsonia intracellularis induces rapid induction of humoral immunity, reduction of bacterial shedding and provides robust gut barrier function. Vaccine 36:1500\u20131508. https:\/\/doi.org\/10.1016\/j.vaccine.2017.12.049","journal-title":"Vaccine"},{"key":"3786_CR31","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2018.00678","volume":"9","author":"ME Schmidt","year":"2018","unstructured":"Schmidt ME, Varga SM (2018) The CD8 T cell response to respiratory virus infections. Front Immunol 9:678. https:\/\/doi.org\/10.3389\/fimmu.2018.00678","journal-title":"Front Immunol"},{"key":"3786_CR32","doi-asserted-by":"publisher","first-page":"109","DOI":"10.1111\/j.1469-0691.2012.03939.x","volume":"18","author":"KL Seib","year":"2012","unstructured":"Seib KL, Zhao X, Rappuoli R (2012) Developing vaccines in the era of genomics: a decade of reverse vaccinology. Clin Microbiol Infect 18:109\u2013116. https:\/\/doi.org\/10.1111\/j.1469-0691.2012.03939.x","journal-title":"Clin Microbiol Infect"},{"key":"3786_CR33","doi-asserted-by":"publisher","first-page":"842","DOI":"10.1039\/C5SC03892H","volume":"7","author":"M Skwarczynski","year":"2016","unstructured":"Skwarczynski M, Toth I (2016) Peptide-based synthetic vaccines. Chem Sci 7:842\u2013854. https:\/\/doi.org\/10.1039\/C5SC03892H","journal-title":"Chem Sci"},{"key":"3786_CR34","doi-asserted-by":"publisher","first-page":"405","DOI":"10.1016\/j.jbi.2014.11.003","volume":"53","author":"RE Soria-Guerra","year":"2015","unstructured":"Soria-Guerra RE, Nieto-Gomez R, Govea-Alonso DO, Rosales-Mendoza S (2015) An overview of bioinformatics tools for epitope prediction: implications on vaccine development. J Biomed Inform 53:405\u2013414. https:\/\/doi.org\/10.1016\/j.jbi.2014.11.003","journal-title":"J Biomed Inform"},{"key":"3786_CR35","doi-asserted-by":"publisher","first-page":"465","DOI":"10.1177\/0300985813520249","volume":"51","author":"FA Vannucci","year":"2014","unstructured":"Vannucci FA, Gebhart CJ (2014) Recent advances in understanding the pathogenesis of Lawsonia intracellularis infections. Vet Pathol 51:465\u2013477. https:\/\/doi.org\/10.1177\/0300985813520249","journal-title":"Vet Pathol"},{"key":"3786_CR36","doi-asserted-by":"publisher","first-page":"1070","DOI":"10.1128\/JCM.05976-11","volume":"50","author":"FA Vannucci","year":"2012","unstructured":"Vannucci FA, Wattanaphansak S, Gebhart CJ (2012) An alternative method for cultivation of Lawsonia intracellularis. J Clin Microbiol 50:1070\u20131072. https:\/\/doi.org\/10.1128\/JCM.05976-11","journal-title":"J Clin Microbiol"},{"key":"3786_CR37","doi-asserted-by":"publisher","first-page":"5535","DOI":"10.1128\/JVI.78.11.5535-5545.2004","volume":"78","author":"EJ Wherry","year":"2004","unstructured":"Wherry EJ, Ahmed R (2004) Memory CD8 T-cell differentiation during viral infection. J Virol 78:5535\u20135545. https:\/\/doi.org\/10.1128\/JVI.78.11.5535-5545.2004","journal-title":"J Virol"},{"key":"3786_CR38","doi-asserted-by":"publisher","first-page":"235","DOI":"10.3390\/biom4010235","volume":"4","author":"H Yamaguchi","year":"2014","unstructured":"Yamaguchi H, Miyazaki M (2014) Refolding techniques for recovering biologically active recombinant proteins from inclusion bodies. Biomolecules 4:235\u2013251. https:\/\/doi.org\/10.3390\/biom4010235","journal-title":"Biomolecules"},{"key":"3786_CR39","doi-asserted-by":"publisher","first-page":"W174","DOI":"10.1093\/nar\/gkv342","volume":"43","author":"J Yang","year":"2015","unstructured":"Yang J, Zhang Y (2015) I-TASSER server: new development for protein structure and function predictions. Nucleic Acids Res 43:W174\u2013W181. https:\/\/doi.org\/10.1093\/nar\/gkv342","journal-title":"Nucleic Acids Res"},{"key":"3786_CR40","doi-asserted-by":"publisher","first-page":"7","DOI":"10.1038\/nmeth.3213","volume":"12","author":"J Yang","year":"2015","unstructured":"Yang J, Yan R, Roy A, Xu D, Poisson J, Zhang Y (2015) The I-TASSER suite: protein structure and function prediction. Nat Methods 12:7\u20138. https:\/\/doi.org\/10.1038\/nmeth.3213","journal-title":"Nat Methods"}],"container-title":["Biotechnology Letters"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1007\/s10529-026-03786-6.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/article\/10.1007\/s10529-026-03786-6","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1007\/s10529-026-03786-6.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T06:20:38Z","timestamp":1789712438000},"score":1,"resource":{"primary":{"URL":"https:\/\/link.springer.com\/10.1007\/s10529-026-03786-6"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,18]]},"references-count":40,"journal-issue":{"issue":"5","published-print":{"date-parts":[[2026,10]]}},"alternative-id":["3786"],"URL":"https:\/\/doi.org\/10.1007\/s10529-026-03786-6","relation":{},"ISSN":["0141-5492","1573-6776"],"issn-type":[{"value":"0141-5492","type":"print"},{"value":"1573-6776","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,18]]},"assertion":[{"value":"28 May 2026","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"14 August 2026","order":2,"name":"revised","label":"Revised","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"2 September 2026","order":3,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"18 September 2026","order":4,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare no conflicts of interest.","order":1,"name":"Ethics","label":"Conflict of interest","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"Schematic representation of pET-28a(+) plasmid constructs encoding chimeric proteins derived from Lawsonia intracellularis. (A) Construct carrying chimera 1 (Li_1). (B) Construct carrying chimera 2 (Li_2). (C) Construct carrying chimera 3 (Li_3). (D) Construct carrying the full-length chimera (Li_full). Numbers indicate nucleotide positions. Key elements, including the T7 promoter, His-tag, and restriction enzyme sites, are indicated.","order":2,"name":"Ethics","label":"Supplementary material","group":{"name":"EthicsHeading","label":"Declarations"}}],"article-number":"115"}}