DNA Vaccines, 1st ed. 2021 Methods and Protocols Methods in Molecular Biology Series, Vol. 2197

This volume details practical procedures on the latest DNA vaccine technology. Chapters guide readers through methods and protocols on DNA vaccine design, the adjuvant influence, production and purification methodologies, delivery systems, and approaches of the influence of DNA vaccines in the immunological response performance and in the cancer immunotherapy. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and cutting-edge, DNA Vaccines: Methods and Protocols aims to ensure successful results in the further study of this vital field.

Part I: Vector design For vaccination

1. A Novel Cre Recombinase Mediated In Vivo Minicircle (CRIM) DNA Vaccine Platform for Veterinary Application

Yanlong Jiang, Guilian Yang, and Chunfeng Wang

2. Vaccination with Messenger RNA – a Promising Alternative to DNA Vaccination

István Tombácz, Drew Weissman, and Norbert Pardi

3. Impact of a Plasmid DNA-Based Alphavirus Vaccine on Immunization Efficiency

Kenneth Lundstrom

Part II: DNA Vaccine Adjuvants and Immunostimulatory Response

4. CpG Oligonucleotides as Vaccine Adjuvants

Neslihan Kayraklioglu, Begum Horuluoglu, and Dennis M. Klinman

5. Molecular Adjuvants for DNA Vaccines: Application, Design, Preparation and Formulation

Ailar Sabbaghi and Amir Ghaemi

6. Assessing Antigen-specific Cellular Immune Responses Upon HIV/SIV Plasmid DNA Vaccination in the Non-human Primate Model

Part III          Biotechnological Processes to Obtain DNA Vaccines

Luís A. Passarinha

8. Primary Purification of Plasmid DNA Using Differential Isopropanol Precipitation

Alexandra Wagner, A. Rita Silva-Santos, Sara Sousa Rosa, Sophie Gierak, Ana M. Azevedo, and Duarte Miguel F. Prazeres

9. Scale-Up of Plasmid DNA Downstream Process Based on Chromatographic Monoliths

Urh Černigoj and Aleš Štrancar

10. Purification of Plasmid DNA by Multimodal Chromatography

A. Rita Silva-Santos, Sara Sousa Rosa, Duarte Miguel F. Prazeres, and Ana M. Azevedo

11. Minicircle DNA Vaccine Purification and E7 Antigen Expression Assessment

Ana M. Almeida, Dalinda Eusébio, João A. Queiroz, Fani Sousa, and Ângela Sousa

Part IV:         DNA Vaccine Delivery

12. Tumor-Specific CD8⁺ T Cell Responses Induced by DNA Vaccination

13. Therapeutic DNA Vaccine Against HPV16-Associated Cancer

14. Bulk and Microfluidic Synthesis of Stealth and Cationic-Liposomes for Gene Delivery Applications

15. Conception of Plasmid DNA and Polyethylenimine Delivery Systems with Potential Application in DNA Vaccines Field

Diana Costa, Ângela Sousa, Rúben Faria, Ana Raquel Neves, and João A. Queiroz

16. Main Features of DNA Based Vectors for Use in Lactic Acid Bacteria and Update Protocols

Nina D. Coelho-Rocha, Fernanda A. L. Barroso, Laísa M. Tavares, Ester S. S. dos Santos, Vasco Azevedo, Mariana M. Drumond, and Pamela Mancha-Agresti

17. Ethics of DNA Vaccines Transfer for Clinical Research

Ana Cristina Ramalhinhoand Miguel Castelo-Branco

18. Preparation of an Academic Clinical TrialPreparation of an academic clinical trial

Ana Cristina Ramalhinhoand Miguel Castelo-Branco

Includes cutting-edge methods and protocols

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from the experts