Development of a biotechnological process for the production of recombinant canine interferon-alpha using the baculovirus-insect cell and larvae system.

IF 2.1 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biotechnology Letters Pub Date : 2025-09-11 DOI:10.1007/s10529-025-03644-x

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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引用次数: 0

Abstract

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.

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利用杆状病毒-昆虫细胞和幼虫系统生产重组犬α干扰素的生物工艺研究。

目前，拉丁美洲对兽医产品的需求正在大幅增加，以确保动物健康。特别是干扰素（ifn）是一种细胞因子，在小动物临床治疗病毒性或肿瘤性疾病中起着重要作用。我们开发了一种利用杆状病毒-昆虫细胞和幼虫系统生产重组犬干扰素CaIFN-α7 （rCaIFN-α7）的生物技术工艺，并评估了其抗病毒和抗肿瘤的生物活性，并与市场上可用的兽医干扰素猫干扰素（rFeIFN-ω, Virbagen omega®）进行了比较。通过固定化金属离子色谱法一步纯化Sf9细胞中表达的重组rCaIFN-α7，产量为4.43 mg/l，纯度为95%。纯化的rCaIFN-α7对犬狼疮肾（ATCC NBL-2）感染水疱性口炎病毒（VSV，新泽西株）的犬肾细胞株MDCK （NBL-2， Madin Darby犬肾）具有抗病毒活性，纯化后的回收率为5.34 × 107 AU/ml。此外，rCaIFN-α7对犬粘膜黑色素瘤细胞株Ak和Bk具有抗肿瘤活性。在典型的放大工艺中，用200只拉氏鼠幼体获得了具有生物活性的rCaIFN-α7，其抗病毒效价为4.5 × 107 AU/ml，产率为5.36 × 106 AU/g。该产品对犬粘膜黑色素瘤细胞系Mc也显示出抗肿瘤活性，并且从起始样品中纯度很高，但产率为20%，可能是由于聚集或降解事件导致生物活性降低。

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来源期刊

Biotechnology Letters 工程技术-生物工程与应用微生物

CiteScore

5.90

自引率

3.70%

发文量

108

审稿时长

1.2 months

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.