Evaluation of eco-friendly zwitterionic detergents for enveloped virus inactivation
Evaluation of eco-friendly zwitterionic detergents for enveloped virus inactivation
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DOI：
10.1002/bit.26209
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发表时间：
2017-04-01
影响因子：
3.8
通讯作者：
Ghose, Sanchayita
中科院分区：
文献类型：
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作者：
Conley, Lynn;Tao, Yinying;Ghose, Sanchayita
在蛋白质生物治疗纯化过程中加入去污剂是一种简单且非常可靠的灭活包膜病毒的方法。去垢剂 Triton X-100 已使用多年，是多种商业治疗性蛋白质生产过程的一部分。然而，最近的生态研究表明，Triton X-100 及其分解产物可能对水生生物产生内分泌干扰物，从环境影响的角度引发担忧。因此，一些司法管辖区对 Triton X-100 向废水处理厂的排放进行了监管，并且需要用于病毒灭活的替代洗涤剂。在这项工作中，我们报告了对作为 Triton X-100 可行替代品的更环保清洁剂的识别和评估。最初鉴定了五种对环境影响低至中度的候选洗涤剂，并对其蛋白质稳定性进行了评估，然后使用包膜病毒模型进行了概念验证病毒灭活研究。从一组候选物中，十二烷基二甲胺 N-氧化物 (LDAO) 被认为是最有前途的洗涤剂，因为它具有低生态毒性、强大的抗病毒活性（在 X-MuLX 验证设定点条件下，LRV > 4），并且对蛋白质功能没有任何负面影响。该去垢剂在多种蛋白质浓度、溶液电导率、pH 值以及多种不同的细胞培养液基质中表现出有效且强大的病毒灭活作用。与病毒灭活效力降低相关的唯一工艺参数是 LDAO 浓度，并且仅当浓度降低至低于洗涤剂的临界胶束浓度 (CMC) 时。此外，这项工作还证明，在 Protein A 亲和层析后，LDAO 被清除至低于可检测水平，使其适合在利用这种层析模式进行蛋白质捕获的平台过程中使用。所有这些发现表明，LDAO 可能是 Triton X-100 的实用替代品，用于灭活包膜病毒的蛋白质治疗生产过程。生物技术。生物工程。 2017；114：813-820。 (c) 2016 年 Wiley 期刊公司。
Inclusion of a detergent in protein biotherapeutic purification processes is a simple and very robust method for inactivating enveloped viruses. The detergent Triton X-100 has been used for many years and is part of the production process of several commercial therapeutic proteins. However, recent ecological studies have suggested that Triton X-100 and its break-down products can potentially behave as endocrine disrupters in aquatic organisms, raising concerns from an environmental impact perspective. As such, discharge of Triton X-100 into the waste water treatment plants is regulated in some jurisdictions, and alternative detergents for viral inactivation are required. In this work, we report on the identification and evaluation of more eco-friendly detergents as viable replacements for Triton X-100. Five detergent candidates with low to moderate environmental impact were initially identified and evaluated with respect to protein stability, followed by proof-of-concept virus inactivation studies using a model enveloped virus. From the set of candidates lauryldimethylamine N-oxide (LDAO) was identified as the most promising detergent due to its low ecotoxicity, robust anti-viral activity (LRV >4 at validation set-point conditions with X-MuLX), and absence of any negative impact on protein function. This detergent exhibited effective and robust virus inactivation in a broad range of protein concentrations, solution conductivities, pHs, and in several different cell culture fluid matrices. The only process parameter which correlated with reduced virus inactivation potency was LDAO concentration, and then only when the concentration was reduced to below the detergent's critical micelle concentration (CMC). Additionally, this work also demonstrated that LDAO was cleared to below detectable levels after Protein A affinity chromatography, making it suitable for use in a platform process that utilizes this chromatographic mode for protein capture. All these findings suggest that LDAO may be a practical alternative to Triton X-100 for use in protein therapeutic production processes for inactivating enveloped viruses. Biotechnol. Bioeng. 2017;114: 813-820. (c) 2016 Wiley Periodicals, Inc.