Exploring Elastin-Like Polypeptide Tags and Mini-intein for Recombinant Protein Purification in Leishmania tarentolae
Recombinant technology has enabled the production of a wide range of proteins with applications in various fields, including reproduction, food supplements, and medicine. However, the high costs associated with producing and purifying recombinant proteins limit their widespread use in the veterinary...
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| Format: | article |
| Sprog: | engelsk |
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2026
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| Online adgang: | https://hdl.handle.net/20.500.12381/5457 https://doi.org/10.1016/j.pep.2026.106902 |
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| _version_ | 1868889995323899904 |
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| author | Bassetti, Lucía |
| author2 | Crispo, Martina Bollati-Fogolín, Mariela Pantano, Sergio Comini, Marcelo A. Abreu, Cecilia |
| author2_role | author author author author author |
| author_browse | Abreu, Cecilia Bassetti, Lucía Bollati-Fogolín, Mariela Comini, Marcelo A. Crispo, Martina Pantano, Sergio |
| author_facet | Bassetti, Lucía Crispo, Martina Bollati-Fogolín, Mariela Pantano, Sergio Comini, Marcelo A. Abreu, Cecilia |
| author_role | author |
| collection | IPMON en REDI |
| dc.creator.none.fl_str_mv | Bassetti, Lucía Crispo, Martina Bollati-Fogolín, Mariela Pantano, Sergio Comini, Marcelo A. Abreu, Cecilia |
| dc.date.none.fl_str_mv | 2026-03-06T19:32:58Z 2026-03-06T19:32:58Z 2026-02-16 |
| dc.identifier.none.fl_str_mv | https://hdl.handle.net/20.500.12381/5457 FMV_3_2018_1_148443 POS_NAC_2020_1_164355 https://doi.org/10.1016/j.pep.2026.106902 |
| dc.language.none.fl_str_mv | eng |
| dc.publisher.none.fl_str_mv | Elsevier |
| dc.rights.none.fl_str_mv | Acceso abierto info:eu-repo/semantics/openAccess Reconocimiento-NoComercial-SinObraDerivada 4.0 Internacional. (CC BY-NC-ND) |
| dc.source.none.fl_str_mv | Protein Expression and Purification reponame:IPMON en REDI instname:Institut Pasteur de Montevideo instacron:Institut Pasteur de Montevideo |
| dc.subject.none.fl_str_mv | Protein expression and purification Elastin-Like Polypeptide (ELP) Leishmania tarentolae mini-intein ΔI-CM Ciencias Naturales y Exactas Ciencias Biológicas Bioquímica y Biología Molecular Biología Celular, Microbiología |
| dc.title.none.fl_str_mv | Exploring Elastin-Like Polypeptide Tags and Mini-intein for Recombinant Protein Purification in Leishmania tarentolae |
| dc.type.none.fl_str_mv | Artículo info:eu-repo/semantics/article Aceptado info:eu-repo/semantics/acceptedVersion |
| description | Recombinant technology has enabled the production of a wide range of proteins with applications in various fields, including reproduction, food supplements, and medicine. However, the high costs associated with producing and purifying recombinant proteins limit their widespread use in the veterinary field. Therefore, the development of alternative, low-cost strategies for production and purification is essential to address these challenges. The non-pathogenic eukaryotic protozoan Leishmania tarentolae has emerged as an affordable expression system for heterologous proteins, providing glycosylation profiles similar to those of higher eukaryotes. Despite this, the cost of recombinant protein purification remains a significant barrier to expanding the use of recombinant products in areas such as livestock. In this study, we evaluate an unconventional purification tag, the elastin-like polypeptide (ELP), for purifying recombinant proteins secreted by L. tarentolae. We also explored the use of the mini-intein ΔI-CM as a cleavable linker between ELP and the target protein. This intein was found to undergo intracellular self-cleavage, precluding its use in secretion-based applications in this host. Despite this limitation, the ELP-mediated purification from the culture supernatant was successfully achieved with a high purity using 0.6 M ammonium sulfate at 25-30 °C. To our knowledge, this is the first report of ELP-based protein purification in a protozoan host, representing a promising tool with broad potential for applications in molecular biology, pharmaceutical development, and other fields that require high-purity proteins with an improved cost-to-benefit ratio. |
| eu_rights_str_mv | openAccess |
| format | article |
| id | anni_2c833ea4a343bd998707fcb9e682ef60 |
| identifier_str_mv | FMV_3_2018_1_148443 POS_NAC_2020_1_164355 |
| instacron_str | Institut Pasteur de Montevideo |
| institution | Institut Pasteur de Montevideo |
| instname_str | Institut Pasteur de Montevideo |
| language | eng |
| network_acronym_str | anni |
| network_name_str | oai-lr-anni |
| oai_identifier_str | oai:redi.anii.org.uy:20.500.12381/5457 |
| publishDate | 2026 |
| publishDateSort | 2026 |
| publisher.none.fl_str_mv | Elsevier |
| reponame_str | IPMON en REDI |
| repository.mail.fl_str_mv | |
| repository.name.fl_str_mv | |
| repository_id_str | |
| rights_invalid_str_mv | Acceso abierto Reconocimiento-NoComercial-SinObraDerivada 4.0 Internacional. (CC BY-NC-ND) |
| spelling | Exploring Elastin-Like Polypeptide Tags and Mini-intein for Recombinant Protein Purification in Leishmania tarentolaeBassetti, LucíaCrispo, MartinaBollati-Fogolín, MarielaPantano, SergioComini, Marcelo A.Abreu, CeciliaProtein expression and purificationElastin-Like Polypeptide (ELP)Leishmania tarentolaemini-intein ΔI-CMCiencias Naturales y ExactasCiencias BiológicasBioquímica y Biología MolecularBiología Celular, MicrobiologíaRecombinant technology has enabled the production of a wide range of proteins with applications in various fields, including reproduction, food supplements, and medicine. However, the high costs associated with producing and purifying recombinant proteins limit their widespread use in the veterinary field. Therefore, the development of alternative, low-cost strategies for production and purification is essential to address these challenges. The non-pathogenic eukaryotic protozoan Leishmania tarentolae has emerged as an affordable expression system for heterologous proteins, providing glycosylation profiles similar to those of higher eukaryotes. Despite this, the cost of recombinant protein purification remains a significant barrier to expanding the use of recombinant products in areas such as livestock. In this study, we evaluate an unconventional purification tag, the elastin-like polypeptide (ELP), for purifying recombinant proteins secreted by L. tarentolae. We also explored the use of the mini-intein ΔI-CM as a cleavable linker between ELP and the target protein. This intein was found to undergo intracellular self-cleavage, precluding its use in secretion-based applications in this host. Despite this limitation, the ELP-mediated purification from the culture supernatant was successfully achieved with a high purity using 0.6 M ammonium sulfate at 25-30 °C. To our knowledge, this is the first report of ELP-based protein purification in a protozoan host, representing a promising tool with broad potential for applications in molecular biology, pharmaceutical development, and other fields that require high-purity proteins with an improved cost-to-benefit ratio.Agencia Nacional de Investigación e InnovaciónFondo para la Convergencia Estructural del MercosurElsevier2026-03-06T19:32:58Z2026-03-06T19:32:58Z2026-02-16Artículoinfo:eu-repo/semantics/articleAceptadoinfo:eu-repo/semantics/acceptedVersionhttps://hdl.handle.net/20.500.12381/5457FMV_3_2018_1_148443POS_NAC_2020_1_164355https://doi.org/10.1016/j.pep.2026.106902Protein Expression and Purificationreponame:IPMON en REDIinstname:Institut Pasteur de Montevideoinstacron:Institut Pasteur de MontevideoengAcceso abiertoinfo:eu-repo/semantics/openAccessReconocimiento-NoComercial-SinObraDerivada 4.0 Internacional. (CC BY-NC-ND)oai:redi.anii.org.uy:20.500.12381/54572026-06-16T05:20:22Z |
| spellingShingle | Exploring Elastin-Like Polypeptide Tags and Mini-intein for Recombinant Protein Purification in Leishmania tarentolae Bassetti, Lucía Protein expression and purification Elastin-Like Polypeptide (ELP) Leishmania tarentolae mini-intein ΔI-CM Ciencias Naturales y Exactas Ciencias Biológicas Bioquímica y Biología Molecular Biología Celular, Microbiología |
| status_str | acceptedVersion |
| title | Exploring Elastin-Like Polypeptide Tags and Mini-intein for Recombinant Protein Purification in Leishmania tarentolae |
| title_full | Exploring Elastin-Like Polypeptide Tags and Mini-intein for Recombinant Protein Purification in Leishmania tarentolae |
| title_fullStr | Exploring Elastin-Like Polypeptide Tags and Mini-intein for Recombinant Protein Purification in Leishmania tarentolae |
| title_full_unstemmed | Exploring Elastin-Like Polypeptide Tags and Mini-intein for Recombinant Protein Purification in Leishmania tarentolae |
| title_short | Exploring Elastin-Like Polypeptide Tags and Mini-intein for Recombinant Protein Purification in Leishmania tarentolae |
| title_sort | Exploring Elastin-Like Polypeptide Tags and Mini-intein for Recombinant Protein Purification in Leishmania tarentolae |
| topic | Protein expression and purification Elastin-Like Polypeptide (ELP) Leishmania tarentolae mini-intein ΔI-CM Ciencias Naturales y Exactas Ciencias Biológicas Bioquímica y Biología Molecular Biología Celular, Microbiología |
| url | https://hdl.handle.net/20.500.12381/5457 https://doi.org/10.1016/j.pep.2026.106902 |