{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,6]],"date-time":"2025-12-06T05:30:22Z","timestamp":1764999022098,"version":"3.46.0"},"reference-count":45,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2025,12,4]],"date-time":"2025-12-04T00:00:00Z","timestamp":1764806400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"FIOCRUZ\/CNPq","award":["PAEF II","IOC-008-FIO-22-2-49"],"award-info":[{"award-number":["PAEF II","IOC-008-FIO-22-2-49"]}]},{"DOI":"10.13039\/501100004586","name":"Faperj","doi-asserted-by":"crossref","award":["E-26\/211-707\/2021","E-26\/202.854\/2019","E-26\/202.760\/2019","E26\/211.112\/2019"],"award-info":[{"award-number":["E-26\/211-707\/2021","E-26\/202.854\/2019","E-26\/202.760\/2019","E26\/211.112\/2019"]}],"id":[{"id":"10.13039\/501100004586","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Antibodies"],"abstract":"<jats:p>Backgroud\/Objectives: Canine Visceral Leishmaniasis (CVL), caused by Leishmania infantum, is a significant public health concern due to dogs serving as reservoirs for human infection. An accurate and rapid diagnostic method to distinguish symptomatic and asymptomatic CVL from healthy and vaccinated animals is essential for controlling canine and human disease. Developing innovative antibody detection techniques and exploring new antigens are essential for enhancing CVL testing efficiency. Our study focuses on a multiplex flow cytometry technique to detect Leishmania-specific antibodies in canine serum. This involved conjugating small peptides with carrier proteins or peptide tags, sequences designed to facilitate bead coupling. Methods: A peptide from the L. infantum A2 protein was coupled to beads in three forms: unconjugated, conjugated with BSA, and conjugated with a C-terminal \u03b2-alanine\u2013lysine (x4)\u2013cysteine TAG. This TAG was previously designed to enhance peptide solubility, improve binding efficiency, and provide functional groups for covalent attachment to the beads, ensuring stable immobilization in the multiplex assay. Results: Our results suggest that the multiplex approach shows promise as a rapid serological test for CVL, particularly with TAG-conjugated peptides, which optimize bead coupling. However, peptide\/BSA conjugation revealed anti-BSA antibodies in samples from healthy and CVL dogs. Conclusions: In conclusion, our findings highlight the potential of multiplex methodologies to enhance CVL diagnostics and caution against using BSA as a bead coupling agent in serological tests for canine samples due to its impact on test specificity and sensitivity.<\/jats:p>","DOI":"10.3390\/antib14040103","type":"journal-article","created":{"date-parts":[[2025,12,4]],"date-time":"2025-12-04T13:53:30Z","timestamp":1764856410000},"page":"103","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Comparison of Antigen Conjugation to a Peptidic Carrier or to Bovine Serum Albumin in the Serodiagnosis of Canine Visceral Leishmaniasis via Suspension Array Technology"],"prefix":"10.3390","volume":"14","author":[{"given":"Thais Stelzer","family":"Toledo","sequence":"first","affiliation":[{"name":"Laboratory of Immunoparasitology, Oswaldo Cruz Institute (IOC), Funda\u00e7\u00e3o Oswaldo Cruz (Fiocruz), Rio de Janeiro 21040-900, Brazil"},{"name":"Postgraduate Program in Parasitic Biology, Oswaldo Cruz Institute (IOC), Funda\u00e7\u00e3o Oswaldo Cruz (Fiocruz), Rio de Janeiro 21040-900, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Pauline Martins","family":"Cunha","sequence":"additional","affiliation":[{"name":"Laboratory of Immunoparasitology, Oswaldo Cruz Institute (IOC), Funda\u00e7\u00e3o Oswaldo Cruz (Fiocruz), Rio de Janeiro 21040-900, Brazil"},{"name":"Professional Master\u2019s in Immunobiological Technology, Immunobiological Technology Institute (Bio-Manguinhos), Funda\u00e7\u00e3o Oswaldo Cruz (Fiocruz), Rio de Janeiro 21040-900, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5848-404X","authenticated-orcid":false,"given":"Josu\u00e9 da Costa","family":"Lima-Junior","sequence":"additional","affiliation":[{"name":"Laboratory of Immunoparasitology, Oswaldo Cruz Institute (IOC), Funda\u00e7\u00e3o Oswaldo Cruz (Fiocruz), Rio de Janeiro 21040-900, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Monique Paiva","family":"De Campos","sequence":"additional","affiliation":[{"name":"Leishmaniasis Reference Laboratory, Carlos Chagas Institute (ICC), Funda\u00e7\u00e3o Oswaldo Cruz (Fiocruz), Curitiba 81350-010, Brazil"},{"name":"Postgraduate Program in Clinical Research in Infectious Diseases, Instituto Nacional de Infectologia Evandro Chagas (INI), Funda\u00e7\u00e3o Oswaldo Cruz (Fiocruz), Rio de Janeiro 21040-900, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alinne R. S.","family":"Renzetti","sequence":"additional","affiliation":[{"name":"Laboratory of Immunoparasitology, Oswaldo Cruz Institute (IOC), Funda\u00e7\u00e3o Oswaldo Cruz (Fiocruz), Rio de Janeiro 21040-900, Brazil"},{"name":"Postgraduate Program in Clinical Research in Infectious Diseases, Instituto Nacional de Infectologia Evandro Chagas (INI), Funda\u00e7\u00e3o Oswaldo Cruz (Fiocruz), Rio de Janeiro 21040-900, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fabiano Borges","family":"Figueiredo","sequence":"additional","affiliation":[{"name":"Postgraduate Program in Clinical Research in Infectious Diseases, Instituto Nacional de Infectologia Evandro Chagas (INI), Funda\u00e7\u00e3o Oswaldo Cruz (Fiocruz), Rio de Janeiro 21040-900, Brazil"},{"name":"Cellular Biology Laboratory, Carlos Chagas Institute (ICC), Funda\u00e7\u00e3o Oswaldo Cruz (Fiocruz), Curitiba 81350-010, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7188-251X","authenticated-orcid":false,"given":"Fernanda Nazar\u00e9","family":"Morgado","sequence":"additional","affiliation":[{"name":"Laboratory of Immunoparasitology, Oswaldo Cruz Institute (IOC), Funda\u00e7\u00e3o Oswaldo Cruz (Fiocruz), Rio de Janeiro 21040-900, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Renato","family":"Porrozzi","sequence":"additional","affiliation":[{"name":"Protozoology Laboratory, Oswaldo Cruz Institute (IOC), Funda\u00e7\u00e3o Oswaldo Cruz (Fiocruz), Rio de Janeiro 21040-900, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0114-5001","authenticated-orcid":false,"given":"Fatima da","family":"Concei\u00e7\u00e3o-Silva","sequence":"additional","affiliation":[{"name":"Laboratory of Immunoparasitology, Oswaldo Cruz Institute (IOC), Funda\u00e7\u00e3o Oswaldo Cruz (Fiocruz), Rio de Janeiro 21040-900, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marta de Almeida","family":"Santiago","sequence":"additional","affiliation":[{"name":"Laboratory of Immunoparasitology, Oswaldo Cruz Institute (IOC), Funda\u00e7\u00e3o Oswaldo Cruz (Fiocruz), Rio de Janeiro 21040-900, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2017-8602","authenticated-orcid":false,"given":"Paula Mello","family":"De Luca","sequence":"additional","affiliation":[{"name":"Laboratory of Immunoparasitology, Oswaldo Cruz Institute (IOC), Funda\u00e7\u00e3o Oswaldo Cruz (Fiocruz), Rio de Janeiro 21040-900, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,12,4]]},"reference":[{"key":"ref_1","unstructured":"Salomon, O.D. 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