7. Either inactivated virus or recombinant viral proteins can be
used but the concentration of antigen might need to be optimized for each antigen.
8. Bang the plate onto absorbent pads to remove air bubbles and
excess moisture in between every two consecutive washes.
9. To maintain appropriate cell sensitivity, Vero cells should be in
the log-growth phase and therefore cultured for a minimum of
48 h prior to being harvested for cell plate preparation.
10. Typically, cell numbers should usually range from 2 Â 10
5 to
4 Â 10
5 cells per well but might need to be optimized.
11. Do not stack the plates if you have more than one to prevent
edge effects and do not move the plates while culturing.
Acknowledgments
This work was supported by National Institute of Allergy and
Infectious Diseases of the National Institutes of Health under
Contracts
No.
HHS-N272200800039C
and
HHS-N272201400053C.
References
1. Li L, Petrovsky N (2017) Molecular adjuvants
for DNA vaccines. Curr Issues Mol Biol
22:17–40
2. Li L, Saade F, Petrovsky N (2012) The future
of human DNA vaccines. J Biotechnol 162
(2–3):171–182
3. Porter KR et al (2011) Immunogenicity and
protective efficacy of a vaxfectin-adjuvanted
tetravalent dengue DNA vaccine. Vaccine 30
(2):336–341
4. Nukuzuma C et al (2003) Enhancing effect of
vaxfectin on the ability of a Japanese encephalitis DNA vaccine to induce neutralizing antibody in mice. Viral Immunol 16(2):183–189
5. Ara Y et al (2001) Zymosan enhances the
immune response to DNA vaccine for human
immunodeficiency virus type-1 through the
activation of complement system. Immunology
103(1):98–105
6. Honda-Okubo Y, Saade F, Petrovsky N (2012)
Advax, a polysaccharide adjuvant derived from
delta inulin, provides improved influenza vaccine protection through broad-based enhancement of adaptive immune responses. Vaccine
30(36):5373–5381
7. Lobigs M et al (2010) An inactivated Vero cellgrown
Japanese
encephalitis
vaccine
formulated with Advax, a novel inulin-based
adjuvant, induces protective neutralizing antibody against homologous and heterologous
flaviviruses. J Gen Virol 91(Pt 6):1407–1417
8. Cukjati D et al (2007) Real time electroporation control for accurate and safe in vivo
non-viral gene therapy. Bioelectrochemistry
70(2):501–507
9. Becker SM, Kuznetsov AV (2007) Local temperature rises influence in vivo electroporation
pore development: a numerical stratum corneum lipid phase transition model. J Biomech
Eng 129(5):712–721
10. Wu CJ et al (2004) In vivo electroporation of
skeletal muscles increases the efficacy of Japanese encephalitis virus DNA vaccine. Vaccine
22(11–12):1457–1464
11. Livingston BD et al (2009) Comparative performance of a licensed anthrax vaccine versus
electroporation based delivery of a PA encoding DNA vaccine in rhesus macaques. Vaccine
28(4):1056–1061
12. Luckay A et al (2007) Effect of plasmid DNA
vaccine design and in vivo electroporation on
the
resulting
vaccine-specific
immune
responses in rhesus macaques. J Virol 81
(10):5257–5269
DNA Vaccine Design and Adjuvant Selection
421
used but the concentration of antigen might need to be optimized for each antigen.
8. Bang the plate onto absorbent pads to remove air bubbles and
excess moisture in between every two consecutive washes.
9. To maintain appropriate cell sensitivity, Vero cells should be in
the log-growth phase and therefore cultured for a minimum of
48 h prior to being harvested for cell plate preparation.
10. Typically, cell numbers should usually range from 2 Â 10
5 to
4 Â 10
5 cells per well but might need to be optimized.
11. Do not stack the plates if you have more than one to prevent
edge effects and do not move the plates while culturing.
Acknowledgments
This work was supported by National Institute of Allergy and
Infectious Diseases of the National Institutes of Health under
Contracts
No.
HHS-N272200800039C
and
HHS-N272201400053C.
References
1. Li L, Petrovsky N (2017) Molecular adjuvants
for DNA vaccines. Curr Issues Mol Biol
22:17–40
2. Li L, Saade F, Petrovsky N (2012) The future
of human DNA vaccines. J Biotechnol 162
(2–3):171–182
3. Porter KR et al (2011) Immunogenicity and
protective efficacy of a vaxfectin-adjuvanted
tetravalent dengue DNA vaccine. Vaccine 30
(2):336–341
4. Nukuzuma C et al (2003) Enhancing effect of
vaxfectin on the ability of a Japanese encephalitis DNA vaccine to induce neutralizing antibody in mice. Viral Immunol 16(2):183–189
5. Ara Y et al (2001) Zymosan enhances the
immune response to DNA vaccine for human
immunodeficiency virus type-1 through the
activation of complement system. Immunology
103(1):98–105
6. Honda-Okubo Y, Saade F, Petrovsky N (2012)
Advax, a polysaccharide adjuvant derived from
delta inulin, provides improved influenza vaccine protection through broad-based enhancement of adaptive immune responses. Vaccine
30(36):5373–5381
7. Lobigs M et al (2010) An inactivated Vero cellgrown
Japanese
encephalitis
vaccine
formulated with Advax, a novel inulin-based
adjuvant, induces protective neutralizing antibody against homologous and heterologous
flaviviruses. J Gen Virol 91(Pt 6):1407–1417
8. Cukjati D et al (2007) Real time electroporation control for accurate and safe in vivo
non-viral gene therapy. Bioelectrochemistry
70(2):501–507
9. Becker SM, Kuznetsov AV (2007) Local temperature rises influence in vivo electroporation
pore development: a numerical stratum corneum lipid phase transition model. J Biomech
Eng 129(5):712–721
10. Wu CJ et al (2004) In vivo electroporation of
skeletal muscles increases the efficacy of Japanese encephalitis virus DNA vaccine. Vaccine
22(11–12):1457–1464
11. Livingston BD et al (2009) Comparative performance of a licensed anthrax vaccine versus
electroporation based delivery of a PA encoding DNA vaccine in rhesus macaques. Vaccine
28(4):1056–1061
12. Luckay A et al (2007) Effect of plasmid DNA
vaccine design and in vivo electroporation on
the
resulting
vaccine-specific
immune
responses in rhesus macaques. J Virol 81
(10):5257–5269
DNA Vaccine Design and Adjuvant Selection
421