Disassembly mediated multimodal chromatography based purification of HPV-VLPs produced in Pichia pastoris

IF 2.2 4区医学 Q3 BIOCHEMICAL RESEARCH METHODS

Journal of virological methods Pub Date : 2025-04-16 DOI:10.1016/j.jviromet.2025.115168

Rashmi Sharma , Pragya Prakash , Lukas Gerstweiler , Anurag S. Rathore

引用次数: 0

Abstract

Human Papillomavirus Virus-Like Particles (HPV-VLPs) are a highly effective vaccine to prevent cervical cancer. Current production and purification processes for HPV-VLPs suffer from poor yield and suboptimal process economics. The current study presents a purification strategy based multi-modal cation exchange chromatography (Capto™ MMC) for the purification of HPV-VLPs produced in Pichia pastoris. Single step purification of disassembled VLPs offered a superior product recovery (> 80 %) and purity (> 70 %) compared to traditional VLP purification platforms that comprise anion exchange and cation exchange chromatography (yield: 32 %, purity: 52 %). Furthermore, it was observed that disassembling the intact VLPs to capsomere subunits before purification provided an improved dynamic binding capacity of up to 18.1 mg/mL (at 2 min residence time), 4 times higher than that with intact HPV-VLPs.

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毕赤酵母中HPV-VLPs的分离纯化

人乳头瘤病毒样颗粒（hpv - vlp）是一种非常有效的预防宫颈癌的疫苗。目前HPV-VLPs的生产和纯化工艺存在产率低和工艺经济不理想的问题。目前的研究提出了一种基于多模态阳离子交换色谱（Capto™MMC）的纯化策略，用于纯化毕赤酵母生产的HPV-VLPs。拆分的VLPs的单步纯化提供了优越的产品回收率(>；80 %)和纯度(>；70 %)，与传统的VLP净化平台相比，包括阴离子交换和阳离子交换色谱（产率：32 %，纯度：52 %）。此外，我们还观察到，在纯化前将完整的VLPs分解到衣壳体亚基上，可以提供高达18.1 mg/mL的动态结合能力（在2 分钟的停留时间），比完整的HPV-VLPs高4倍。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of virological methods 医学-病毒学

CiteScore

5.80

自引率

0.00%

发文量

209

审稿时长

41 days

期刊介绍： The Journal of Virological Methods focuses on original, high quality research papers that describe novel and comprehensively tested methods which enhance human, animal, plant, bacterial or environmental virology and prions research and discovery. The methods may include, but not limited to, the study of: Viral components and morphology- Virus isolation, propagation and development of viral vectors- Viral pathogenesis, oncogenesis, vaccines and antivirals- Virus replication, host-pathogen interactions and responses- Virus transmission, prevention, control and treatment- Viral metagenomics and virome- Virus ecology, adaption and evolution- Applied virology such as nanotechnology- Viral diagnosis with novelty and comprehensive evaluation. We seek articles, systematic reviews, meta-analyses and laboratory protocols that include comprehensive technical details with statistical confirmations that provide validations against current best practice, international standards or quality assurance programs and which advance knowledge in virology leading to improved medical, veterinary or agricultural practices and management.