Maria S Rodriguez, Ignacio Smith, Marcela S Villaverde, Joaquin M Birenbaum, Joaquin Poodts, Federico J Wolman, Alexandra M Targovnik, Maria V Miranda
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Recombinant rCaIFN-α7 expressed in Sf9 cell culture was purified in a single step by immobilized metal ion chromatography, yielding 4.43 mg/l with 95% purity. The purified rCaIFN-α7 showed antiviral activity against the canine kidney cell line MDCK (NBL-2, Madin Darby canine kidney) derived from Canis lupus familiaris kidney (ATCC NBL-2) infected with Vesicular Stomatitis Virus (VSV, New Jersey strain), with a recovery of 5.34 × 10<sup>7</sup> AU/ml after purification. Furthermore, rCaIFN-α7 exhibited antitumor activity against canine mucosal melanoma cell lines Ak and Bk. In a typical scale-up process using 200 Rachiplusia nu larvae, biologically active rCaIFN-α7 was obtained, showing an antiviral titer of 4.5 × 10<sup>7</sup> AU/ml and a yield of 5.36 × 10<sup>6</sup> AU/g of larvae. This product also showed antitumor activity against canine mucosal melanoma cell line Mc and a high degree of purity from the starting sample, but a yield of 20%, probably due to aggregation or degradation events leading to a decrease in biological activity.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 5","pages":"101"},"PeriodicalIF":2.1000,"publicationDate":"2025-09-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Development of a biotechnological process for the production of recombinant canine interferon-alpha using the baculovirus-insect cell and larvae system.\",\"authors\":\"Maria S Rodriguez, Ignacio Smith, Marcela S Villaverde, Joaquin M Birenbaum, Joaquin Poodts, Federico J Wolman, Alexandra M Targovnik, Maria V Miranda\",\"doi\":\"10.1007/s10529-025-03644-x\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Currently, Latin America is experiencing a considerable increase in demand for veterinary products to ensure animal health. In particular, interferons (IFNs) are cytokines that play an important role in small animal clinics for the treatment of viral or oncological diseases. We developed a biotechnological process for the production of recombinant canine interferon CaIFN-α7 (rCaIFN-α7) using the baculovirus-insect cell and larvae system, and evaluated its biological activity, both antiviral and antitumor, for comparison with the veterinary interferon available on the market, feline omega interferon (rFeIFN-ω, Virbagen Omega®). Recombinant rCaIFN-α7 expressed in Sf9 cell culture was purified in a single step by immobilized metal ion chromatography, yielding 4.43 mg/l with 95% purity. The purified rCaIFN-α7 showed antiviral activity against the canine kidney cell line MDCK (NBL-2, Madin Darby canine kidney) derived from Canis lupus familiaris kidney (ATCC NBL-2) infected with Vesicular Stomatitis Virus (VSV, New Jersey strain), with a recovery of 5.34 × 10<sup>7</sup> AU/ml after purification. Furthermore, rCaIFN-α7 exhibited antitumor activity against canine mucosal melanoma cell lines Ak and Bk. In a typical scale-up process using 200 Rachiplusia nu larvae, biologically active rCaIFN-α7 was obtained, showing an antiviral titer of 4.5 × 10<sup>7</sup> AU/ml and a yield of 5.36 × 10<sup>6</sup> AU/g of larvae. 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Development of a biotechnological process for the production of recombinant canine interferon-alpha using the baculovirus-insect cell and larvae system.
Currently, Latin America is experiencing a considerable increase in demand for veterinary products to ensure animal health. In particular, interferons (IFNs) are cytokines that play an important role in small animal clinics for the treatment of viral or oncological diseases. We developed a biotechnological process for the production of recombinant canine interferon CaIFN-α7 (rCaIFN-α7) using the baculovirus-insect cell and larvae system, and evaluated its biological activity, both antiviral and antitumor, for comparison with the veterinary interferon available on the market, feline omega interferon (rFeIFN-ω, Virbagen Omega®). Recombinant rCaIFN-α7 expressed in Sf9 cell culture was purified in a single step by immobilized metal ion chromatography, yielding 4.43 mg/l with 95% purity. The purified rCaIFN-α7 showed antiviral activity against the canine kidney cell line MDCK (NBL-2, Madin Darby canine kidney) derived from Canis lupus familiaris kidney (ATCC NBL-2) infected with Vesicular Stomatitis Virus (VSV, New Jersey strain), with a recovery of 5.34 × 107 AU/ml after purification. Furthermore, rCaIFN-α7 exhibited antitumor activity against canine mucosal melanoma cell lines Ak and Bk. In a typical scale-up process using 200 Rachiplusia nu larvae, biologically active rCaIFN-α7 was obtained, showing an antiviral titer of 4.5 × 107 AU/ml and a yield of 5.36 × 106 AU/g of larvae. This product also showed antitumor activity against canine mucosal melanoma cell line Mc and a high degree of purity from the starting sample, but a yield of 20%, probably due to aggregation or degradation events leading to a decrease in biological activity.
期刊介绍：
Biotechnology Letters is the world’s leading rapid-publication primary journal dedicated to biotechnology as a whole – that is to topics relating to actual or potential applications of biological reactions affected by microbial, plant or animal cells and biocatalysts derived from them.
All relevant aspects of molecular biology, genetics and cell biochemistry, of process and reactor design, of pre- and post-treatment steps, and of manufacturing or service operations are therefore included.
Contributions from industrial and academic laboratories are equally welcome. We also welcome contributions covering biotechnological aspects of regenerative medicine and biomaterials and also cancer biotechnology. Criteria for the acceptance of papers relate to our aim of publishing useful and informative results that will be of value to other workers in related fields.
The emphasis is very much on novelty and immediacy in order to justify rapid publication of authors’ results. It should be noted, however, that we do not normally publish papers (but this is not absolute) that deal with unidentified consortia of microorganisms (e.g. as in activated sludge) as these results may not be easily reproducible in other laboratories.
Papers describing the isolation and identification of microorganisms are not regarded as appropriate but such information can be appended as supporting information to a paper. Papers dealing with simple process development are usually considered to lack sufficient novelty or interest to warrant publication.
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