Characterisation and separation of infectious bursal disease virus-like particles using aqueous two-phase systems

doi:10.1016/j.cjche.2022.08.019

Chinese Journal of Chemical Engineering ›› 2023, Vol. 57 ›› Issue (5): 72-78.DOI: 10.1016/j.cjche.2022.08.019

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Characterisation and separation of infectious bursal disease virus-like particles using aqueous two-phase systems

Hui Yi Leong¹, Xiao-Qian Fu¹, Xiang-Yu Liu², Shan-Jing Yao¹, Dong-Qiang Lin¹

1. Zhejiang Key Laboratory of Smart Biomaterials, Key Laboratory of Biomass Chemical Engineering of Ministry of Education, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China;
2. Sinopharm Animal Health Corporation Ltd., Wuhan 430075, China

Received:2022-07-06 Revised:2022-08-16 Online:2023-07-08 Published:2023-05-28
Contact: Dong-Qiang Lin,E-mail:lindq@zju.edu.cn
Supported by:
The authors would like to thank Zhejiang University and Talent-Introduction Program of China for Postdoctoral Researcher for the financial support. This work was financially supported by the National Key Research & Development Program of China (2021YFE0113300); and the National Natural Science Foundation of China (22078286).

Characterisation and separation of infectious bursal disease virus-like particles using aqueous two-phase systems

Hui Yi Leong¹, Xiao-Qian Fu¹, Xiang-Yu Liu², Shan-Jing Yao¹, Dong-Qiang Lin¹

1. Zhejiang Key Laboratory of Smart Biomaterials, Key Laboratory of Biomass Chemical Engineering of Ministry of Education, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China;
2. Sinopharm Animal Health Corporation Ltd., Wuhan 430075, China

通讯作者: Dong-Qiang Lin,E-mail:lindq@zju.edu.cn
基金资助:
The authors would like to thank Zhejiang University and Talent-Introduction Program of China for Postdoctoral Researcher for the financial support. This work was financially supported by the National Key Research & Development Program of China (2021YFE0113300); and the National Natural Science Foundation of China (22078286).

Abstract

Abstract: Infectious bursal disease (IBD) causes considerable economic losses in the commercial poultry industry worldwide. The principal way to control IBD virus (IBDV), the causative agent of IBD, is still through vaccination programs. Virus-like particles (VLPs) are recognised as a safe and potent recombinant vaccine platform. This research work explores the characterisation and separation of infectious bursal disease virus-like particles (IBD-VLPs) from crude feedstock. Various characteristics were studied with high-performance size-exclusion chromatography (HP-SEC), sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS-PAGE) and transmission electron microscopy (TEM) analyses. Subsequently, the separation of IBD-VLPs using polyethylene glycol (PEG)/sodium citrate-based aqueous two-phase systems (ATPSs) was conducted and optimised. Moreover, a scale-up study of the best ATPS constituted of 15% PEG 6000, 11% sodium citrate and 10% crude feedstock was performed to compare the separation performance of IBD-VLPs with and without centrifugation-assisted. The results indicated that the optimised ATPS with centrifugation-assisted for both 5 g and 50 g systems showed good recovery of IBD-VLPs of >97% in the interphase between the PEG-rich top and salt-rich bottom phases. These optimised systems also showed high removal efficiencies of impurities of >95%. The results demonstrated that aqueous two-phase extraction could be a promising technology for efficient VLPs separation.

Key words: Aqueous two-phase extraction, Infectious bursal disease virus, Polymers, Salt, Separation, Virus-like particle

摘要： Infectious bursal disease (IBD) causes considerable economic losses in the commercial poultry industry worldwide. The principal way to control IBD virus (IBDV), the causative agent of IBD, is still through vaccination programs. Virus-like particles (VLPs) are recognised as a safe and potent recombinant vaccine platform. This research work explores the characterisation and separation of infectious bursal disease virus-like particles (IBD-VLPs) from crude feedstock. Various characteristics were studied with high-performance size-exclusion chromatography (HP-SEC), sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS-PAGE) and transmission electron microscopy (TEM) analyses. Subsequently, the separation of IBD-VLPs using polyethylene glycol (PEG)/sodium citrate-based aqueous two-phase systems (ATPSs) was conducted and optimised. Moreover, a scale-up study of the best ATPS constituted of 15% PEG 6000, 11% sodium citrate and 10% crude feedstock was performed to compare the separation performance of IBD-VLPs with and without centrifugation-assisted. The results indicated that the optimised ATPS with centrifugation-assisted for both 5 g and 50 g systems showed good recovery of IBD-VLPs of >97% in the interphase between the PEG-rich top and salt-rich bottom phases. These optimised systems also showed high removal efficiencies of impurities of >95%. The results demonstrated that aqueous two-phase extraction could be a promising technology for efficient VLPs separation.

关键词: Aqueous two-phase extraction, Infectious bursal disease virus, Polymers, Salt, Separation, Virus-like particle

Hui Yi Leong, Xiao-Qian Fu, Xiang-Yu Liu, Shan-Jing Yao, Dong-Qiang Lin. Characterisation and separation of infectious bursal disease virus-like particles using aqueous two-phase systems[J]. Chinese Journal of Chemical Engineering, 2023, 57(5): 72-78.

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Characterisation and separation of infectious bursal disease virus-like particles using aqueous two-phase systems

Characterisation and separation of infectious bursal disease virus-like particles using aqueous two-phase systems

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