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.

引用

页数：24

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