An optimized method for high-titer lentivirus preparations without   ultracentrifugation

doi:10.1038/srep13875

CORC > 北京大学 > 生命科学学院

	An optimized method for high-titer lentivirus preparations without ultracentrifugation
	Jiang, Wei ; Hua, Rui ; Wei, Mengping ; Li, Chenhong ; Qiu, Zilong ; Yang, Xiaofei ; Zhang, Chen
刊名	SCIENTIFIC REPORTS
	2015
关键词	MEDIATED GENE-TRANSFER VECTOR PREPARATIONS IN-VIVO PURIFICATION DELIVERY SYSTEM ULTRAFILTRATION TRANSDUCTION TITRATION PURPOSES
DOI	10.1038/srep13875
英文摘要	Lentiviral technology has proven to be a powerful tool to express exogenous genes in dividing and non-dividing cells. Currently, most protocols for generating high-titer lentivirus require ultracentrifugation, which can be an instrumental barrier for routine operations in a laboratory. In this study, the effect of relative centrifugal force (RCF) on the concentration efficiency of the lentivirus was systematically explored, and it was found that sucrose gradient centrifugation with a relatively low speed (<= 10,000 g) robustly produces a high-titer virus (up to 2 x 10(8) TU/ml). The optimal sucrose concentration is 10%, and the recovery rate of the functional virus is greater than 80%. The infection efficiency of both concentrated and un-concentrated lentivirus decreases rapidly when the viruses are stored at 4 degrees C (tau approximate to 1.3 days) or subjected to multiple freeze-thaw cycles (tau = 1.1 rounds). In summary, we describe an efficient and easy-to-handle protocol for high-titer lentivirus purification.; National Basic Research Program of China [2011CB809102, 2014CB942804, 2014BAI03B01, 2012YQ0302604]; National Science Foundation of China [31222025, 31171025, 31300892]; National Science Foundation of Hubei [2014CFA027]; SCUN [YZZ13002]; National Century Excellent Talents in University of China [NCET-11-0020]; Seeding Grant for Medicine and Life Sciences of Peking University [2014-MB-11]; SCI(E); PubMed; ARTICLE; sunlittlefly@hotmail.com; ch.zhang@pku.edu.cn; 13875; 5
语种	英语
内容类型	期刊论文
源URL	[http://ir.pku.edu.cn/handle/20.500.11897/416344]
专题	生命科学学院
推荐引用方式 GB/T 7714	Jiang, Wei,Hua, Rui,Wei, Mengping,et al. An optimized method for high-titer lentivirus preparations without ultracentrifugation[J]. SCIENTIFIC REPORTS,2015.
APA	Jiang, Wei.,Hua, Rui.,Wei, Mengping.,Li, Chenhong.,Qiu, Zilong.,...&Zhang, Chen.(2015).An optimized method for high-titer lentivirus preparations without ultracentrifugation.SCIENTIFIC REPORTS.
MLA	Jiang, Wei,et al."An optimized method for high-titer lentivirus preparations without ultracentrifugation".SCIENTIFIC REPORTS (2015).