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mRNA 脂质纳米粒疫苗的研制方案及小鼠免疫效力分析。

Protocol for the development of mRNA lipid nanoparticle vaccines and analysis of immunization efficiency in mice.

机构信息

Department of Immunology and Immunotherapy, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Precision Immunology Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Department of Hematology/Oncology, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.

Department of Immunology and Immunotherapy, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Precision Immunology Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Department of Hematology/Oncology, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Icahn Genomics Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.

出版信息

STAR Protoc. 2024 Jun 21;5(2):103087. doi: 10.1016/j.xpro.2024.103087. Epub 2024 May 23.

DOI:10.1016/j.xpro.2024.103087
PMID:38795353
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11144802/
Abstract

Here, we present a protocol for the development of mRNA-loaded lipid nanoparticle (LNP) vaccines for target antigen sequences of interest. We describe key steps required to design and synthesize mRNA constructs, their LNP encapsulation, and mouse immunization. We then detail quality control assays to determine RNA purity, guidelines to measure RNA immunogenicity using in vitro reporter systems, and a technique to evaluate antigen-specific T cell responses following immunization.

摘要

在这里，我们提出了一种用于负载有目标抗原序列的信使 RNA（mRNA）的脂质纳米颗粒（LNP）疫苗的开发方案。我们描述了设计和合成 mRNA 构建体、它们的 LNP 包封以及小鼠免疫所需的关键步骤。然后，我们详细介绍了质量控制测定，以确定 RNA 纯度，使用体外报告系统测量 RNA 免疫原性的指南，以及评估免疫后抗原特异性 T 细胞反应的技术。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fcea/11144802/31454b049ad2/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fcea/11144802/bb708d6960ae/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fcea/11144802/ee0e5df6803d/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fcea/11144802/c313ca9cac53/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fcea/11144802/263308ae800e/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fcea/11144802/31454b049ad2/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fcea/11144802/bb708d6960ae/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fcea/11144802/ee0e5df6803d/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fcea/11144802/c313ca9cac53/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fcea/11144802/263308ae800e/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fcea/11144802/31454b049ad2/gr4.jpg

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