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首页> 外文期刊>Biotechnology Progress >Reduction of the Infectivity of Baculovirus Stocks Frozen at Ultra-Low Temperature in Serum-Free Media: The Role of Lipid Emulsions
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Reduction of the Infectivity of Baculovirus Stocks Frozen at Ultra-Low Temperature in Serum-Free Media: The Role of Lipid Emulsions

机译：在无血清培养基中超低温冷冻的杆状病毒原种的感染性降低：脂质乳剂的作用

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The infectivity of stocks of baculoviruses produced in serum-free media is sensitive to freezing at ultra-low temperatures. The objective of this work was to elucidate the causes of such sensitivity, using as a model the freezing of stocks of Anticarsia gemmatalis multiple nucleopolyhedrovirus (AgMNPV), a baculovirus widely employed as biological insecticide. Titers of supernatants of cell cultures infected with AgMNPV in four different serum-free media supplemented with lipid emulsions were reduced by 50 to 90% after six months freezing. By using a full factorial experiment, freezing and lipid emulsion, as well as the interaction between them, were identified as the main factors reducing the viral titer. The virucidal effect of the lipid emulsion was reproduced by one of their components, the surfactant Polysorbate 80. Damaged viral envelopes were observed by transmission electron microscopy in most particles frozen in a medium supplemented with lipid emulsion or Polysorbate 80. Additionally, Polysorbate 80 also affected the infectivity of AgMNPV stocks that were incubated at 27 degrees C. The identification of the roles played by the lipid emulsion and Polysorbate 80 is not only a contribution to the understanding of the mechanisms underlying the inactivation of baculovirus stocks produced in serum-free media during storage at ultra-low temperature, but is also an input for the rational development of new procedures aimed at improving both the preservation of baculovirus stocks and the composition of culture media for the production of baculovirus-based bioproducts in insect cells. (C) 2016 American Institute of Chemical Engineers
机译：在无血清培养基中产生的杆状病毒原种的感染性对在超低温下冷冻敏感。这项工作的目的是通过冻结冷冻抗角虫多核多角体病毒（AgMNPV）（一种广泛用作生物杀虫剂的杆状病毒）的种群作为模型，阐明这种敏感性的原因。冷冻六个月后，在补充了脂质乳剂的四种不同无血清培养基中，被AgMNPV感染的细胞培养上清液的滴度降低了50％至90％。通过全因子实验，冷冻和脂质乳状液以及它们之间的相互作用被确定为降低病毒滴度的主要因素。脂质乳液的杀病毒作用通过它们的一种成分表面活性剂聚山梨酯80再现。通过透射电子显微镜观察，冷冻在添加有脂质乳液或聚山梨酯80的培养基中冷冻的大多数颗粒中的病毒被膜都受到了破坏。此外，聚山梨酯80也受到了影响。在27摄氏度下孵育的AgMNPV菌种的感染性。脂乳剂和Polysorbate 80的作用的鉴定不仅有助于了解无血清培养基中产生的杆状病毒菌种在培养过程中失活的潜在机制。储存在超低温下，但这也是合理开发旨在改善杆状病毒原种的保存和在昆虫细胞中生产基于杆状病毒的生物制品的培养基组成的新程序的投入。 （C）2016美国化学工程师学会

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