Microfluidic electrical cell lysis for high-throughput and continuous production of cell-free varicella-zoster virus

• Published in: Journal of biotechnology Vol. 335; pp. 19 - 26
• Main Authors: Won, Eun-Jae, Thai, Duc Anh, Duong, Duong Duy, Lee, Nae Yoon, Song, Yoon-Jae
• Format: Journal Article
• Language: English
• Published: Elsevier B.V 20.07.2021
• Subjects: Cell-free VZV; Electrolysis; Focused electric field; Microfluidics; Focused electric field; Electrolysis; Microfluidics; Cell-free VZV
• ISSN: 0168-1656; 1873-4863
• DOI: 10.1016/j.jbiotec.2021.06.006

•A microfluidic device integrated with arrays of microelectrode was developed to continuously produce cell-free VZV.•Multiple constricted channels and microelectrode arrays were designed to generate a high electrolysis efficiency.•The integrated microfluidic platform accomplished a production yield of 1.4 – 2.1×104 PFU mL–1 of cell-free VZV.•Rapid and high-throughput production of the high-titer cell-free viruses was obtained with relatively low voltage input. Varicella-zoster virus (VZV), the causative agent of varicella and herpes zoster, is highly cell-associated and spreads via cell-to-cell contact in tissue culture. The lack of cell-free VZV hampers studies on VZV biology as well as antiviral and vaccine development. In the present study, a poly(methylmethacrylate) microfluidic device integrated with arrays of microelectrode was fabricated to continuously electrolyse VZV-infected cells to produce cell-free viruses. By designing multiple constrictions and microelectrode arrays, a high electric field is focused on the constricted region of the microchannel to disrupt large numbers of virus-infected cells with high-throughput on a microfluidic platform. Plaque assay and scanning electron microscopy were conducted to quantify and characterize cell-free VZV produced using the microfluidic continuous-flow electrical cell lysis device. The process of microfluidic electrical cell lysis followed by subsequent filtration and virus concentration process yielded a 1.4–2.1 × 104 plaque-forming units (PFUs) per mL of cell-free VZV from 7.0 × 106 VZV-infected human foreskin fibroblasts (HFF) cells. The high electric field formed inside a microfluidic channel combined with the continuous-flow of virus-infected cells within the microchannel enabled the rapid and efficient production of high-titer cell-free virus in large quantities with relatively low input of the voltage.