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Influenza Virus Precision Diagnosis and Continuous Purification Enabled by Neuraminidase-Resistant Glycopolymer-Coated Microbeads

Gefei Li, Wenxiao Ma, Juan Mo, Boyang Cheng, Shin-ichiro Shoda, Demin Zhou*, and Xin-Shan Ye*
Cite this: ACS Appl. Mater. Interfaces 2021, 13, 39, 46260–46269
Publication Date:September 22, 2021

https://doi.org/10.1021/acsami.1c11561

Abstract
Rapid diagnosis and vaccine development are critical to prevent the threat posed by viruses. However, rapid tests, such as colloidal gold assays, yield false-negative results due to the low quantities of viruses; moreover, conventional virus purification, including ultracentrifugation and nanofiltration, is multistep and time-consuming, which limits laboratory research and commercial development of viral vaccines. A rapid virus enrichment and purification technique will improve clinical diagnosis sensitivity and simplify vaccine production. Hence, we developed the surface-glycosylated microbeads (glycobeads) featuring chemically synthetic glycoclusters and reversible linkers to selectively capture the influenza virus. The surface plasmon resonance (SPR) evaluation indicated broad spectrum affinity of S-linked glycosides to various influenza viruses. The magnetic glycobeads were integrated into clinical rapid diagnosis, leading to a 30-fold lower limit of detection. Additionally, the captured viruses can be released under physiological conditions, delivering purified viruses with >50% recovery and without decreasing their native infectivity. Notably, this glycobead platform will facilitate the sensitive detection and continuous one-step purification of the target virus that contributes to future vaccine production.
快速诊断和疫苗开发对于预防病毒构成的威胁至关重要。然而，由于病毒数量少，快速检测（如胶体金检测）会产生假阴性结果；此外，包括超速离心和纳滤在内的常规病毒纯化是多步骤且耗时的，这限制了病毒疫苗的实验室研究和商业开发。快速病毒富集和纯化技术将提高临床诊断的敏感性并简化疫苗生产。因此，我们开发了具有化学合成糖簇和可逆接头的表面糖基化微珠（糖珠），以选择性地捕获流感病毒。表面等离子共振 (SPR) 评估表明 S-连接糖苷对各种流感病毒具有广谱亲和力。磁性糖珠被整合到临床快速诊断中，导致检测下限降低 30 倍。此外，捕获的病毒可以在生理条件下释放，以> 50％的回收率提供纯化的病毒，并且不会降低其天然传染性。值得注意的是，这个糖珠平台将促进目标病毒的灵敏检测和连续一步纯化，有助于未来的疫苗生产。

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