Purification of dengue virus like particle from insect cell culture

:53–9 18 Miller N (2010) Recent progress in dengue vaccine research and development Curr Opin Mol Ther 2010 Feb;12(1):31-8 19 Mukhopadhyay S, Kuhn RJ, Rossmann MG (2005) A structural perspective of the flavivirus life cycle Nat Rev Microbiol 2005 Jan; 3(1):13-22 20 Murphy BR, Whitehead SS (2011) Immune response to dengue virus and prospects for a vaccine Annu Rev Immunol 2011; 29: 587–619 21 Normile D Tropical medicine (2013) Surprising new dengue virus throws a spanner in disease control efforts Science 2013;342(6157):415 22 Perera R, Kuhn RJ (2008) Structural proteomics of dengue virus Curr Opin Microbiol 2008; 11:369–77 23 Rodenhuis-Zybert IA, van der Schaar HM, da Silva Voorham JM, van der EndeMetselaar H, Lei HY, Wilschut J, et al (2010) Immature dengue virus: a veiled pathogen PLoS Pathog 2010; 6:e1000718 24 Rogers, D J., Wilson, A J., Hay, S I & Graham, A J (2006) The global distribution of yellow fever and dengue Adv Parasitol 62, 181–220 (2006) 25 Simmons, C P., Farrar, J J., van Vinh Chau, N & Wills, B (2012) Dengue N Engl J Med 366, 1423–1432 (2012) 26 Singhasivanon P1, Jacobson J (2009) Dengue is a major global health problem J Clin Virol 2009 Oct;46 Suppl 2:S1-2 27 Solomonides, Tony (2010) Healthgrid applications and core technologies: proceedings of HealthGrid 2010 ([Online-Ausg.] ed.) Amsterdam: IOS Press p 235 ISBN 978-1-60750-582-2 28 Staropoli I, Frenkiel MP, Megret F, et al (1997) Affinity-purified dengue-2 virus envelope glycoprotein induces neutralizing antibodies and protective immunity in mice Vaccine 1997, 15:1946-54 39 29 Yorgo Modis, Steven Ogata, David Clements, Stephen C Harrison (2004) Structure of the dengue virus envelope protein after membrane fusion Nature 427, 313-319 30 Zhang Y, Zhang W, Ogata S, Clements D, Strauss JH, Baker TS,Kuhn RJ, Rossmann MG (2004) Conformational changes of the flavivirus E glycoprotein Structure 12:1607-1618 31 Zhang, S., Liang, M., Gu, W., Li, C., Miao, F., Wang, X., Jin, C., Zhang, L., Zhang, F., Zhang, Q., Jiang, L., Li, M., Li, D (2011) Vaccination with dengue virus-like particles induces humoral and cellular immune responses in mice Virol J 8, 333 Internet resource Dengue vaccine Initiative (2016) (http://www.denguevaccines.org/live-attenuatedvaccines ) (Accessed 2016) Dengue Virus Net (2016) (http://www.denguevirusnet.com/vaccine-research.html ) (Accessed 2016) Dengue Virus Net (2016) ( http://www.denguevirusnet.com/dengue-virus.html) (Accessed 2016) Nature Education (2014) http://www.nature.com/scitable/content/types-of-denguevirus-vaccines-22405302) (Accessed 2014) UKHealthCentre (2016) (http://www.healthcentre.org.uk/vaccine/types-ofvaccine.html ) (Accessed 27 August 2016) WHO Dengue and dengue haemorrhagic fever Factsheet No 117, revised May 2008 Geneva, World Health Organization, 2008 (http://www.who.int/mediacentre/factsheets/fs117/en/ ) (Acceessed May 2008) WHO Dengue Guidelines for Diagnosis, Treatment, Prevention and Control Geneva: World Health Organization (2009) (http://whqlibdoc.who.int/publications/2009/9789241547871_eng.pdf?ua=1 ) (Accessed Feb 24, 2014) 40 APPENDIX 0.5M Tris-HCl (100ml) • Tris: 6.05g • Dissolve in 70ml distilled water • Adjust the pH to 6.8 with HCl and make up to 100ml • Store at 4°C 1.5M Tris-HCl 100ml) • 18.15g • Dissolve in 70ml distilled water • Adjust the pH to 8.8 with HCl and make up to 100ml • Store at 4°C 10% Amino persulphate (10% APS) (1ml) • Ammonium persulphate: 100mg • Dissolve in 70ml distilled water (prepare fresh everytime) 10% resolving (2gel) • 30% Bis-Acrylamide: 6.075ml • 1.5M Tris-HCl (pH=8.8): 4.95ml • 10% SDS: 150àl ã DI water: 3.75ml ã TEMED: 7.5 àl • 10% APS: 75 µl 10% SDS (100ml) • SDS: 10g • O : 90ml 41 12% resolving • 30% Bis-Acrylamide: 5.94ml • 1.5M Tris-HCl (pH=8.8): 3.75ml • 10% SDS: 150àl ã DI water: 5.085ml ã TEMED: 7.5 µl • 10% APS: 75 µl 4% stocking • 30% Bis-Acrylamide: 0.99ml • 1.5M Tris-HCl (pH=8.8): 1.89ml • 10% SDS: 75àl ã DI water: 4.5ml ã TEMED: 7.5 àl • 10% APS: 37.5 µl Binding buffer (1L) Make 20mM sodium phosphate • 0.2M di-sodium phosphate ( ) (100ml) di-sodium phosphate : 3.56g Dissolve in 100ml distilled water • 0.2M O (100ml) mono-sodium phosphate : 2.76g Dissolve in 100ml distilled water • Adjust pH by mixing mono-sodium phosphate and di-sodium phosphate • Mix 100ml Sodium phosphate and 900ml distilled water 42 Blocking buffer 3% BSA (50ml) • BSA: 1.5g • Add 30ml PBST • Mix well up to 50ml 10 Blocking buffer 5% skim milk (50 ml) • Skim milk: 2.5g • Add PBST and mix well up to 50ml 11 Destanining Solution (500ml) • Methanol: 200ml • Acetic acid: 35ml • Up to 500ml with distilled water 12 Developeing buffer (5ml) (AP conjuagate substrate kit) • Development buffer: 200ml • Reagent A: 50µl • Reagent B: 50àl ã DI water: 4.7ml 13 Developing solution (100ml) • 500µl of 1% citric (0.01g/1ml DI) • 50µl 37% formaldehyde • Up to 100ml with DI water 14 Elution buffer (1L) Make 20mM sodium phosphate • 0.2M di-sodium phosphate ( di-sodium phosphate : 3.56g Dissolve in 100ml distilled water 43 ) (100ml) • 0.2M O (100ml) mono-sodium phosphate : 2.76g Dissolve in 100ml distilled water • Adjust pH by mixing mono-sodium phosphate and di-sodium phosphate • Mix 100ml Sodium phosphate and 900ml distilled water Mix 100ml Sodium phosphate with 2M NaCl and up to 1L with distilled water 15 PBS 10X (250ml, pH=7.4) • NaCl: 20g • KCl: 0.5g • : 0.6g • : 3.6g • Dissolve in 100ml distilled water and adjust pH by NaOH up to 7.4 16 PBST (500ml) • 1X PBS: 497.5ml • 0.5% Tween 20: 2.5ml 17 Running buffer 10X (500ml, pH=8.3) • 250mM Tris: 15.15g • 1.92M glycerin: 72g • 1% SDS: 2.5g • Dissolve in distilled water and up to 500ml 18 Silver nitrate solution • 0.8g Ag / 4ml DI water • 0.36% NaOH: 21ml • OH: 1.4ml • Up to 100ml with DI water 44 19 Staining solution (500ml) • Commassie blue: 0.125g • Methanol: 200ml • Acetic acid: 35ml • Adjust volume by distilled water 20 Transfer buffer (500ml) • Methanol: 100ml • Tris base: 7.21g • Glycine: 1.52g • Up to 500ml with distilled water 45 ... (Accessed 2016) Dengue Virus Net (2016) (http://www.denguevirusnet.com/vaccine-research.html ) (Accessed 2016) Dengue Virus Net (2016) ( http://www.denguevirusnet.com /dengue- virus. html) (Accessed... Vaccination with dengue virus- like particles induces humoral and cellular immune responses in mice Virol J 8, 333 Internet resource Dengue vaccine Initiative (2016) (http://www.denguevaccines.org/live-attenuatedvaccines... http://www.nature.com/scitable/content/types -of- denguevirus-vaccines-22405302) (Accessed 2014) UKHealthCentre (2016) (http://www.healthcentre.org.uk/vaccine/types-ofvaccine.html ) (Accessed 27 August 2016) WHO Dengue and dengue haemorrhagic