An optimized measles virus glycoprotein-pseudotyped lentiviral vector production system to promote efficient transduction of human primary B cells

被引：4
|
作者：
Vamva, Eirini [1 ,2 ,3 ]
Ozog, Stosh [1 ,3 ]
Verhoeyen, Els [4 ]
James, Richard G. [2 ,3 ]
Rawlings, David J. [2 ,3 ]
Torbett, Bruce E. [1 ,2 ,3 ,5 ]
机构：
[1] Scripps Res Inst, Dept Immunol & Microbiol, La Jolla, CA 92037 USA
[2] Seattle Childrens Res Inst, Ctr Immun & Immunotherapies, Seattle, WA 98121 USA
[3] Univ Washington, Sch Med, Dept Pediat, Seattle, WA 98195 USA
[4] Univ Lyon, CIRI Int Ctr Infectiol Res, Team EVIR, Lyon, France
[5] Inst Stem Cell & Regenerat Med, Seattle, WA 98109 USA
来源：
STAR PROTOCOLS | 2022年 / 3卷 / 01期
关键词：
Biotechnology and bioengineering; Cell isolation; Flow Cytometry/Mass Cytometry; Immunology; Microbiology; Molecular Biology;
D O I：
10.1016/j.xpro.2022.101228
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
Measles virus envelope pseudotyped LV (MV-LV) can achieve high B cell trans-duction rates (up to 50%), but suffers from low titers. To overcome current limi-tations, we developed an optimized MV-LV production protocol that achieved consistent B cell transduction efficiency up to 75%. We detail this protocol along with analytical assays to assess the results of MV-LV mediated B cell transduction, including flow cytometry for B cell phenotypic characterization and measurement of transduction efficiency, and ddPCR for VCN analysis.
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页数：24
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