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RNA Vaccines

Overview of attention for book
Cover of 'RNA Vaccines'

Table of Contents

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    Book Overview
  2. Altmetric Badge
    Chapter 1 Introduction to RNA Vaccines.
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    Chapter 2 Self-Replicating RNA.
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    Chapter 3 Self-Replicating RNA Vaccine Delivery to Dendritic Cells.
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    Chapter 4 Plant Expression of Trans-Encapsidated Viral Nanoparticle Vaccines with Animal RNA Replicons.
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    Chapter 5 RNActive® Technology: Generation and Testing of Stable and Immunogenic mRNA Vaccines.
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    Chapter 6 Nucleoside Modified mRNA Vaccines for Infectious Diseases.
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    Chapter 7 Generation and Evaluation of Prophylactic mRNA Vaccines Against Allergy.
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    Chapter 8 Measuring the Adjuvant Activity of RNA Vaccines.
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    Chapter 9 Generation of Immunostimulating 130 nm Protamine-RNA nanoparticles.
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    Chapter 10 Electroporation of mRNA as Universal Technology Platform to Transfect a Variety of Primary Cells with Antigens and Functional Proteins.
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    Chapter 11 Adjuvant-Enhanced mRNA Vaccines.
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    Chapter 12 Enhanced Delivery of DNA or RNA Vaccines by Electroporation.
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    Chapter 13 The European Regulatory Environment of RNA-Based Vaccines.
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    Chapter 14 Discovery and Subtyping of Neo-Epitope Specific T-Cell Responses for Cancer Immunotherapy: Addressing the Mutanome.
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    Chapter 15 Considerations for Producing mRNA Vaccines for Clinical Trials.
  17. Altmetric Badge
    Chapter 16 Nonclinical Safety Testing of RNA Vaccines.
  18. Altmetric Badge
    Chapter 17 Immunotherapy of Uveal Melanoma: Vaccination Against Cancer.
Attention for Chapter 10: Electroporation of mRNA as Universal Technology Platform to Transfect a Variety of Primary Cells with Antigens and Functional Proteins.
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Chapter title
Electroporation of mRNA as Universal Technology Platform to Transfect a Variety of Primary Cells with Antigens and Functional Proteins.
Chapter number 10
Book title
RNA Vaccines
Published in
Methods in molecular biology, January 2017
DOI 10.1007/978-1-4939-6481-9_10
Pubmed ID
Book ISBNs
978-1-4939-6479-6, 978-1-4939-6481-9
Authors

Kerstin F. Gerer, Stefanie Hoyer, Jan Dörrie, Niels Schaft, Gerer, Kerstin F., Hoyer, Stefanie, Dörrie, Jan, Schaft, Niels

Editors

Thomas Kramps, Knut Elbers

Abstract

Electroporation (EP) of mRNA into human cells is a broadly applicable method to transiently express proteins of choice in a variety of different cell types. We have spent more than a decade to optimize and adapt this method, first for antigen-loading of dendritic cells (DCs), and subsequently for T cells, B cells, bulk PBMCs, and several cell lines. In this regard, antigens were introduced, processed, and presented in context of MHC class I and II. Next to that, functional proteins like adhesion receptors, T-cell receptors (TCRs), chimeric antigen receptors (CARs), constitutively active signal transducers, and others were successfully expressed. We have also established this protocol under full GMP compliance as part of a manufacturing license to produce mRNA-electroporated DCs for therapeutic vaccination in clinical trials. Therefore, we here want to share our universal mRNA electroporation protocol and the experience we have gathered with this method. The advantages of the transfection method presented here are: (1) easy adaptation to different cell types, (2) scalability from 10(6) to approximately 10(8) cells per shot, (3) high transfection efficiency (80-99 %), (4) homogenous protein expression, (5) GMP compliance if the EP is performed in a class A clean room, and (6) no transgene integration into the genome. The provided protocol involves: Opti-MEM® as EP medium, a square-wave pulse with 500 V, and 4 mm cuvettes. To adapt the protocol to differently sized cells, simply the pulse time is altered. Next to the basic protocol, we also provide an extensive list of hints and tricks, which in our opinion are of great value for everyone who intends to use this transfection technique.

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The data shown below were collected from the profile of 1 X user who shared this research output. Click here to find out more about how the information was compiled.
Mendeley readers

Mendeley readers

The data shown below were compiled from readership statistics for 58 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 58 100%

Demographic breakdown

Readers by professional status Count As %
Researcher 8 14%
Student > Master 8 14%
Student > Bachelor 5 9%
Student > Ph. D. Student 5 9%
Other 3 5%
Other 6 10%
Unknown 23 40%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 9 16%
Agricultural and Biological Sciences 7 12%
Medicine and Dentistry 6 10%
Immunology and Microbiology 4 7%
Pharmacology, Toxicology and Pharmaceutical Science 4 7%
Other 2 3%
Unknown 26 45%
Attention Score in Context

Attention Score in Context

This research output has an Altmetric Attention Score of 1. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 12 January 2018.
All research outputs
#15,409,382
of 22,919,505 outputs
Outputs from Methods in molecular biology
#5,360
of 13,132 outputs
Outputs of similar age
#256,467
of 420,541 outputs
Outputs of similar age from Methods in molecular biology
#466
of 1,074 outputs
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So far Altmetric has tracked 13,132 research outputs from this source. They receive a mean Attention Score of 3.4. This one is in the 44th percentile – i.e., 44% of its peers scored the same or lower than it.
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