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首页> 外文期刊>Nepal Journal of Biotechnology >Effect of lyophilization on infectivity and viral load of Adenovirus
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Effect of lyophilization on infectivity and viral load of Adenovirus

机译：冻干对腺病毒感染性和病毒载量的影响

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Freeze drying (Lyophilization) performed at temperature and pressure below the triple point is being practiced for the preservation of virus stocks for longer periods. The present study is aimed to lyophilize adenovirus strain to study its effects on infectivity and viral load. In-house adenovirus reference strain (stock virus) was propagated in Hep-2 cell line in 25cm2 cell culture flasks. In 24-well plates the serial dilutions of stock virus from 10-1 to 10-7 (100μl inoculum) was inoculated in each well with Hep-2 cells for TCID50 titer and viral DNA was extracted separately to determine viral load by Taqman Real Time PCR. Stock virus was lyophilized in 3 lots and stored at RT (25±2°C) and 4°C separately for 1, 4 and 6 months and subjected to TCID50 (for viral infectivity) and viral load assay (for total viral genome copies). Following lyophilisation and storage of adenoviral strains at RT and 4°C separately did not affect significantly on the viral stability, infectivity as well as viral copy number till 4 months. However, storage at RT for 6 months resulted in 1 log reduction in viral copy number. Thus, storage of even lyophilized virus stock would necessitate a temperature of at least 4°C for prolonged periods. The present study could successfully lyophilize adenovirus and retain its infectivity over a period of 6 months when stored at RT and 4°C. No significant difference in the infectivity or TCID50 titer was observed in the lyophilized virus as compared to the stock virus. However, the viral load was observed to increase with lyophilization of the virus over 6 months when stored at 4°C which possibly is due to the concentration of the virus on freeze-drying.Nepal Journal of Biotechnology. Dec. 2015 Vol. 3, No. 1: 15-21
机译：正在实践在低于三点的温度和压力下进行冷冻干燥（冻干），以更长时间地保存病毒原种。本研究旨在冻干腺病毒株，以研究其对感染性和病毒载量的影响。在25cm2细胞培养瓶中的Hep-2细胞系中繁殖内部腺病毒参考菌株（原始病毒）。在24孔板中，将每个病毒从10-1到10-7的系列稀释液（100μl接种物）接种到每个孔中，用Hep-2细胞接种TCID50滴度，并通过Taqman Real Time分别提取病毒DNA以确定病毒载量PCR。将储备病毒冷冻干燥3批，分别在室温（25±2°C）和4°C下保存1、4和6个月，并进行TCID50（用于病毒感染性）和病毒载量测定（用于总病毒基因组拷贝） 。冷冻干燥和分别在室温和4°C下保存腺病毒株后，对病毒稳定性，感染性和病毒拷贝数直至4个月均无明显影响。然而，在室温下保存6个月导致病毒拷贝数减少了1 log。因此，即使冻干的病毒原种也需要长时间保持至少4°C的温度。本研究可以成功地冻干腺病毒，并在室温和4°C下保存6个月以上。与原种病毒相比，在冻干病毒中未观察到传染性或TCID50滴度的显着差异。然而，在4°C下保存6个月后，观察到病毒载量随病毒冻干而增加，这可能是由于冷冻干燥时病毒的浓度所致。尼泊尔生物技术杂志。 2015年12月3号1：15-21

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