Improving stability of virus-like particles by ion-exchange chromatographic supports with large pore size: Advantages of gigaporous media beyond enhanced binding capacity

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	Improving stability of virus-like particles by ion-exchange chromatographic supports with large pore size: Advantages of gigaporous media beyond enhanced binding capacity
	Yu, Mengran 1,2,3; Li, Yan 1,2; Zhang, Songping 1,2; Li, Xiunan 1,2; Yang, Yanli 1,2,3; Chen, Yi 1,2,3; Ma, Guanghui 1,2; Su, Zhiguo 1,2
刊名	JOURNAL OF CHROMATOGRAPHY A
	2014-02-28
卷号	1331 期号:1 页码:69-79
关键词	Pore size effect Ion-exchange chromatography Virus-like particles (VLPs) Gigaporous media Binding capacity Stability
ISSN号	0021-9673
其他题名	J. Chromatogr. A
中文摘要	Limited binding capacity and low recovery of large size multi-subunits virus-like particles (VLPs) in conventional agarose-gel based chromatographic supports with small pores have long been a bottleneck limiting the large scale purification and application of VLPs. In this study, four anion exchange media including DEAE-Sepharose FF(DEAE-FF), DEAE-Capto, gigaporous DEAE-AP-120 nm and DEAE-AP-280 nm with average pore diameters of 32 nm, 20 nm, 120 nm and 280 nm, respectively, were applied for purification of hepatitis B virus surface antigen (HBsAg) VLPs. Pore size effects of media on the VLPs adsorption equilibrium, adsorption kinetics, dynamic binding capacity (DBC), and recovery were investigated in detail. According to the confocal laser scanning microscopy observation, adsorption of the VLPs in DEAE-FF and DEAE-Capto was mostly confined to a thin shell on the outer surface of the beads, leaving the underlying pore space and the binding sites inaccessibly, while the large pores in gigaporous media enabled the VLPs to access to the interior pore spaces by diffusion transport efficiently. Compared to the most widely used DEAE-FF, gigaporous media DEAE-AP-280 nm gained about 12.9 times increase in static adsorption capacity, 8.0 times increase in DBC, and 11.4 times increase in effective pore diffusivity. Beyond increasing the binding capacity and enhancing the mass transfer, the gigaporous structure also significantly improved the stability of the VLPs during intensive adsorption-desorption process by lowing the multi-point interaction between the VLPs and binding sites in the pores. At 2.0 mg/mL-media loading quantity, about 85.5% VLPs were correctly self-assembled after the chromatography with DEAE-AP-280 nm media; oppositely about 85.2% VLPs lost their normal assembly with DEAE-FF due to irreversible disassembly. Comparative investigation was made to study the purifying performance of these four chromatographic media for actual VLPs purification from recombinant Hansenula polymorpha. DEAE-AP-280 nm media were demonstrated the best results showing the highest recovery of 68.33% and purification fold of 3.47, at 2.98 mg protein/mL-media loading quantity and a flow rate of 240 cm/h. (C) 2014 Elsevier B.V. All rights reserved.
英文摘要	Limited binding capacity and low recovery of large size multi-subunits virus-like particles (VLPs) in conventional agarose-gel based chromatographic supports with small pores have long been a bottleneck limiting the large scale purification and application of VLPs. In this study, four anion exchange media including DEAE-Sepharose FF(DEAE-FF), DEAE-Capto, gigaporous DEAE-AP-120 nm and DEAE-AP-280 nm with average pore diameters of 32 nm, 20 nm, 120 nm and 280 nm, respectively, were applied for purification of hepatitis B virus surface antigen (HBsAg) VLPs. Pore size effects of media on the VLPs adsorption equilibrium, adsorption kinetics, dynamic binding capacity (DBC), and recovery were investigated in detail. According to the confocal laser scanning microscopy observation, adsorption of the VLPs in DEAE-FF and DEAE-Capto was mostly confined to a thin shell on the outer surface of the beads, leaving the underlying pore space and the binding sites inaccessibly, while the large pores in gigaporous media enabled the VLPs to access to the interior pore spaces by diffusion transport efficiently. Compared to the most widely used DEAE-FF, gigaporous media DEAE-AP-280 nm gained about 12.9 times increase in static adsorption capacity, 8.0 times increase in DBC, and 11.4 times increase in effective pore diffusivity. Beyond increasing the binding capacity and enhancing the mass transfer, the gigaporous structure also significantly improved the stability of the VLPs during intensive adsorption-desorption process by lowing the multi-point interaction between the VLPs and binding sites in the pores. At 2.0 mg/mL-media loading quantity, about 85.5% VLPs were correctly self-assembled after the chromatography with DEAE-AP-280 nm media; oppositely about 85.2% VLPs lost their normal assembly with DEAE-FF due to irreversible disassembly. Comparative investigation was made to study the purifying performance of these four chromatographic media for actual VLPs purification from recombinant Hansenula polymorpha. DEAE-AP-280 nm media were demonstrated the best results showing the highest recovery of 68.33% and purification fold of 3.47, at 2.98 mg protein/mL-media loading quantity and a flow rate of 240 cm/h. (C) 2014 Elsevier B.V. All rights reserved.
WOS标题词	Science & Technology ; Life Sciences & Biomedicine ; Physical Sciences
类目[WOS]	Biochemical Research Methods ; Chemistry, Analytical
研究领域[WOS]	Biochemistry & Molecular Biology ; Chemistry
关键词[WOS]	B SURFACE-ANTIGEN ; SHORT MONOLITHIC COLUMNS ; GRAFTED AGAROSE MEDIA ; CHINESE-HAMSTER OVARY ; PROTEIN ADSORPTION ; HANSENULA-POLYMORPHA ; POLYSTYRENE MICROSPHERES ; LARGE BIOMOLECULES ; MOSAIC-VIRUS ; PURIFICATION
收录类别	SCI
原文出处	://WOS:000332264900009
语种	英语
WOS记录号	WOS:000332264900009
公开日期	2014-05-06
内容类型	期刊论文
版本	出版稿
源URL	[http://ir.ipe.ac.cn/handle/122111/8124]
专题	过程工程研究所_研究所（批量导入）
作者单位	1.Chinese Acad Sci, Inst Proc Engn, Natl Key Lab Biochem Engn, Beijing 100190, Peoples R China 2.PLA Key Lab Biopharmaceut Proc & Formulat Engn, Beijing 100190, Peoples R China 3.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
推荐引用方式 GB/T 7714	Yu, Mengran,Li, Yan,Zhang, Songping,et al. Improving stability of virus-like particles by ion-exchange chromatographic supports with large pore size: Advantages of gigaporous media beyond enhanced binding capacity[J]. JOURNAL OF CHROMATOGRAPHY A,2014,1331(1):69-79.
APA	Yu, Mengran.,Li, Yan.,Zhang, Songping.,Li, Xiunan.,Yang, Yanli.,...&Su, Zhiguo.(2014).Improving stability of virus-like particles by ion-exchange chromatographic supports with large pore size: Advantages of gigaporous media beyond enhanced binding capacity.JOURNAL OF CHROMATOGRAPHY A,1331(1),69-79.
MLA	Yu, Mengran,et al."Improving stability of virus-like particles by ion-exchange chromatographic supports with large pore size: Advantages of gigaporous media beyond enhanced binding capacity".JOURNAL OF CHROMATOGRAPHY A 1331.1(2014):69-79.