DNA Vaccines Methods and Protocols

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

Xintao Hu, Barbara K. Felber, and Antonio Valentin

 

Part III          Biotechnological Processes to Obtain DNA Vaccines

7. Enhanced Biosynthesis of Plasmid DNA from Escherichia coli Applying Experimental Design

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 Cernigoj 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

Pablo Cáceres-Morgado and Alvaro Lladser

 

13. Therapeutic DNA Vaccine Against HPV16-Associated Cancer

Meihua Yu and Janin Chandra

 

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

Lucimara Gaziola de la Torre, Amanda da Costa e Silva de Noronha Pessoa, Bruna Gregatti de Carvalho, Thiago Bezerra Taketa, Ismail Es, and Gabriel Perli

 

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

 

Part V:           DNA Vaccine Transfer to Clinical Trials

17. Ethics of DNA Vaccines Transfer for Clinical Research

Ana Cristina Ramalhinho and Miguel Castelo-Branco

 

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

Ana Cristina Ramalhinho and Miguel Castelo-Branco