{"title":"Immunoinformatic approaches for ErpY-LemA chimeric protein design for use in leptospirosis control.","authors":"Elsa Giovanna Ávila-Martínez, Thayná Laner Cardoso, Isabel Ladeira Pereira, Pâmela Scaraffuni Caballero, Daniela Rodriguero Wozeak, Amilton Clair Pinto Seixas Neto, Luciano da Silva Pinto, Daiane Drawanz Hartwig","doi":"10.1093/jambio/lxae179","DOIUrl":null,"url":null,"abstract":"<p><strong>Aims: </strong>Currently, immunoinformatic approaches have shown promise in rapidly and cost-effectively identifying new antigens from the Leptospira proteome. Chimeric multiepitope proteins offer a strategy with significant potential for implementation in diagnosis and vaccines development.</p><p><strong>Methods and results: </strong>In this study, we detail the immunoinformatic analyses and design of a new recombinant chimeric protein constructed with epitopes identified from the sequences of ErpY-like and LemA proteins, previously identified as potential antigens for controlling leptospirosis. We expressed the chimeric protein using Escherichia coli heterologous systems, evaluated its antigenicity using serum from naturally infected patients, and its immunogenicity in mice as an animal model, with Freund as an adjuvant. The resulting recombinant chimeric protein, named rErpY-LemA, was successfully expressed and purified using a prokaryotic system, with an expected mass of 35 kDa. Serologic assays using serum samples from naturally infected patients demonstrated recognition of the chimera protein by antibodies present in sera. Animals immunized with the chimera exhibited a significant IgG antibody response from the 7th day (P < 0.001), persisting until day 49 of experimentation, with a titer of 1:12,800 (P < 0.05). Notably, significant production of IgA, IgM, and IgG subclasses was observed in animals immunized with the chimera.</p><p><strong>Conclusions: </strong>These results highlight the promising role of immunoinformatics in rapidly identifying antigens and the potential of chimeric multiepitope proteins in developing effective strategies for leptospirosis control.</p>","PeriodicalId":15036,"journal":{"name":"Journal of Applied Microbiology","volume":null,"pages":null},"PeriodicalIF":3.2000,"publicationDate":"2024-08-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Applied Microbiology","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1093/jambio/lxae179","RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"BIOTECHNOLOGY & APPLIED MICROBIOLOGY","Score":null,"Total":0}
引用次数: 0
Abstract
Aims: Currently, immunoinformatic approaches have shown promise in rapidly and cost-effectively identifying new antigens from the Leptospira proteome. Chimeric multiepitope proteins offer a strategy with significant potential for implementation in diagnosis and vaccines development.
Methods and results: In this study, we detail the immunoinformatic analyses and design of a new recombinant chimeric protein constructed with epitopes identified from the sequences of ErpY-like and LemA proteins, previously identified as potential antigens for controlling leptospirosis. We expressed the chimeric protein using Escherichia coli heterologous systems, evaluated its antigenicity using serum from naturally infected patients, and its immunogenicity in mice as an animal model, with Freund as an adjuvant. The resulting recombinant chimeric protein, named rErpY-LemA, was successfully expressed and purified using a prokaryotic system, with an expected mass of 35 kDa. Serologic assays using serum samples from naturally infected patients demonstrated recognition of the chimera protein by antibodies present in sera. Animals immunized with the chimera exhibited a significant IgG antibody response from the 7th day (P < 0.001), persisting until day 49 of experimentation, with a titer of 1:12,800 (P < 0.05). Notably, significant production of IgA, IgM, and IgG subclasses was observed in animals immunized with the chimera.
Conclusions: These results highlight the promising role of immunoinformatics in rapidly identifying antigens and the potential of chimeric multiepitope proteins in developing effective strategies for leptospirosis control.
期刊介绍:
Journal of & Letters in Applied Microbiology are two of the flagship research journals of the Society for Applied Microbiology (SfAM). For more than 75 years they have been publishing top quality research and reviews in the broad field of applied microbiology. The journals are provided to all SfAM members as well as having a global online readership totalling more than 500,000 downloads per year in more than 200 countries. Submitting authors can expect fast decision and publication times, averaging 33 days to first decision and 34 days from acceptance to online publication. There are no page charges.