Serum Institute of India Pvt. Ltd, Pune 411028, India; Dr. D.Y. Patil Biotechnology and Bioinformatics Institute, Tathawade, Pune 411033, India; Dr. D.Y. Patil Vidyapeeth, Pune 411018, India.
Serum Institute of India Pvt. Ltd, Pune 411028, India.
Vaccine. 2018 Jul 5;36(29):4215-4221. doi: 10.1016/j.vaccine.2018.06.008. Epub 2018 Jun 8.
Foetal Bovine Serum (FBS) and porcine trypsin are one of the essential raw materials used in the manufacturing of cell culture based viral vaccines. Being from animal origin, these raw materials can potentially contaminate the final product by known or unknown adventitious agents. The issue is more serious in case of live attenuated viral vaccines, where there is no inactivation step which can take care of such adventitious agents. It is essential to design production processes which can offer maximum viral clearance potential for animal origin products. Ultraviolet-C irradiation is known to inactivate various adventitious viral agents; however there are limited studies on ultraviolet inactivation of viruses in liquid media. We obtained a recently developed UVivatec ultraviolet-C (UV-C) irradiation based viral clearance system for evaluating its efficacy to inactivate selected model viruses. This system has a unique design with spiral path of liquid allowing maximum exposure to UV-C light of a short wavelength of 254 nm. Five live attenuated vaccine viruses and four other model viruses were spiked in tissue culture media and exposed to UV-C irradiation. The pre and post UV-C irradiation samples were analyzed for virus content to find out the extent of inactivation of various viruses. These experiments showed substantial log reduction for the majority of the viruses with few exceptions based on the characteristics of these viruses. Having known the effect of UV irradiation on protein structure, we also evaluated the post irradiation samples of culture media for growth promoting properties using one of the most fastidious human diploid cells (MRC-5). UV-C exposure did not show any notable impact on the nutritional properties of culture media. The use of an UV-C irradiation based system is considered to be promising approach to mitigate the risk of adventitious agents in cell culture media arising through animal derived products.
胎牛血清(FBS)和猪胰蛋白酶是用于生产基于细胞培养的病毒疫苗的重要原材料之一。由于它们来源于动物,因此这些原材料可能会通过已知或未知的外来污染物污染最终产品。在生产活病毒减毒疫苗的情况下,情况更为严重,因为没有灭活步骤可以处理这些外来污染物。设计能够为动物源产品提供最大病毒清除潜力的生产工艺至关重要。紫外线-C 照射已被证明可以灭活各种外来病毒,但在液体介质中紫外线灭活病毒的研究有限。我们获得了一种最近开发的基于紫外线-C(UV-C)照射的 UVivatec 病毒清除系统,用于评估其灭活选定模型病毒的效果。该系统具有独特的设计,液体采用螺旋路径,可最大限度地暴露于波长为 254nm 的短波紫外线之下。将五种活病毒减毒疫苗和其他四种模型病毒掺入组织培养介质中并进行紫外线-C 照射。对紫外线-C 照射前后的样品进行病毒含量分析,以确定各种病毒的灭活程度。这些实验表明,大多数病毒的对数减少都相当显著,但也有少数例外,这是基于这些病毒的特性。在了解紫外线照射对蛋白质结构的影响之后,我们还使用最挑剔的人类二倍体细胞(MRC-5)评估了培养基中具有生长促进特性的照射后样品。紫外线-C 暴露对培养基的营养特性没有显示出任何明显影响。使用基于紫外线-C 照射的系统被认为是减轻细胞培养介质中源自动物产品的外来污染物风险的有前途的方法。