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优化麻疹病毒糖蛋白假型慢病毒载体生产系统以促进人原代 B 细胞的高效转导。

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

机构信息

Department of Immunology and Microbiology, The Scripps Research Institute, La Jolla, CA, USA.

Center for Immunity and Immunotherapies, Seattle Children's Research Institute, Seattle, WA, USA.

出版信息

STAR Protoc. 2022 Mar 8;3(1):101228. doi: 10.1016/j.xpro.2022.101228. eCollection 2022 Mar 18.

DOI:10.1016/j.xpro.2022.101228
PMID:35284833
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8914380/
Abstract

Measles virus envelope pseudotyped LV (MV-LV) can achieve high B cell transduction rates (up to 50%), but suffers from low titers. To overcome current limitations, 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.

摘要

麻疹病毒包膜假型慢病毒 (MV-LV) 可实现高 B 细胞转导率（高达 50%），但其滴度较低。为了克服当前的限制，我们开发了一种优化的 MV-LV 生产方案，可实现高达 75%的一致 B 细胞转导效率。我们详细介绍了该方案以及分析检测方法，以评估 MV-LV 介导的 B 细胞转导的结果，包括用于 B 细胞表型特征分析和转导效率测量的流式细胞术，以及用于 VCN 分析的 ddPCR。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/487b/8914380/55abaa1a933f/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/487b/8914380/d5d11c54079d/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/487b/8914380/5212322c006d/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/487b/8914380/4b19f1629182/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/487b/8914380/268025322447/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/487b/8914380/cf2b97cab6d8/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/487b/8914380/55abaa1a933f/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/487b/8914380/d5d11c54079d/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/487b/8914380/5212322c006d/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/487b/8914380/4b19f1629182/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/487b/8914380/268025322447/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/487b/8914380/cf2b97cab6d8/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/487b/8914380/55abaa1a933f/gr5.jpg

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