The production of virus-based vaccines in large-scale cultures of animal cells in the early 1950s was the first industrial application of animal cell culture technology. Examples of human vaccines produced in animal cells include the ones against measles, mumps and rubella. Our group has a track record in the development of several human and veterinary virus-based vaccines.

Rotavirus VLP vaccine
Virus like particles constitutes a new vaccine concept. Such particles consist of self-assembled structural proteins from the virus which can elicit an immune response but as they lack the genetic material from the virus are safer vaccines. We have been working on the rational development of a triple-layered virus like particle vaccine against rotavirus using the baculovirus insect cell system as production platform.

HCV retroVLP vaccine
Retrovirus-like particles (retroVLPs) emerged as a new platform of VLPs for the development of vaccines against enveloped virus as hepatitis C (HCV). RetroVLPs possess a lipidic membrane where it is possible to anchor and present protein from HCV. The group is developing these VLPs using a human cell line as production platform.

DNA vaccines
DNA vaccination consists in the injection of genetically engineered DNA rather than a protein or virus based product. Since the efficiency of transfection of naked or complexed DNA is reduced, often viral vehicles are used to deliver the DNA. This is the case of a human adenovirus used in a veterinary DNA vaccine against Peste des Petites Ruminant virus (PPRV) in which the group has been working.

PPRV live- Attenuated vaccine
Whole virus vaccines, either live or inactivated (killed), constitute the vast majority of vaccines in use. Live virus vaccines are prepared from attenuated strains that are almost or completely devoid of pathogenicity but are capable of inducing a protective immune response. Our group has worked on the production and stability during storage of the live attenuated Peste de Petites Ruminant Virus vaccine.

Selected Publications

  1. Sequeira DP, Correia R, Carrondo MJT, Roldao A, Teixeira AP, Alves PM.  2017. Combining stable insect cell lines with baculovirus-mediated expression for multi-HA influenza VLP production. Vaccine. pii: S0264-410X(17)30246-3.  
  2. Sofia B. Carvalho, Mafalda G. Moleirinho, David Wheatley, John Welsh, René Gantier, Paula M. Alves, Cristina Peixoto, Manuel J. T. Carrondo. 2017. Universal label-free In-process Quantification of Influenza Virus-like particles. Biotechnology Journal. 
  3. Sofia B. Carvalho, A. Raquel Fortuna, Michael Wolff, Cristina Peixoto, Paula M. Alves, Udo Reichl, Manuel J.T. Carrondo.  2017. Purification of influenza virus-like particles using sulfated cellulose membrane adsorbers.  Journal of Chemical Technology and Biotechnology.  
  4. Monteiro F, Bernal V, Alves PM. 2017. Combining stable insect cell lines with baculovirus-mediated expression for multi-HA influenza VLP production. BIOTECHNOLOGY AND BIOENGINEERING.  114, 674-684.  
  5. Sequeira DP, Correia R, Carrondo MJT, Roldao A, Teixeira AP, Alves PM.  2017Combining stable insect cell lines with baculovirus-mediated expression for multi-HA influenza VLP production.  Vaccine. pii: S0264-410X(17)30246-3. 
  6. Patrone M, Coroadinha AS, Teixeira AP, Alves PM. 2016Palmitoylation strengthens cholesterol-dependent multimerization and fusion activity of human cytomegalovirus gB. Journal of Biochemical Chemistry. 
  7. Silva AC, Teixeira AP, Alves PM. 2016. Impact of Adenovirus infection in host cell metabolism evaluated by 1H NMR spectroscopy. J Biotechnol. 
  8. Soares HR, Castro R, Tomas HA, Rodrigues AF, Gomes-Alves P, Bellier B, Klatzmann D, Carrondo MJ, Alves PM, Coroadinha AS. 2016. Tetraspanins displayed in retrovirus-derived virus-like particles and their immunogenicity. Vaccine. 
  9. Silva AC, Simão D, Sousa MFQ, Küppers C, Lucas T, Cruz P, Carrondo MJ,  Kochanek S, Alves PM 2015 Human Amniocyte-derived Cells for the Production of Adenoviral Vectors. Biotechnology Journal, May;10(5):760-71
  10. Castro R, Fernandes P, Laske T, Sousa MFQ, Genzel Y, Scharfenberg K, Alves PM, Coroadinha AS. 2015. Production of canine adenovirus type 2 in serum-free suspension cultures of MDCK cells. Appl Microbiol Biotechnol Sep;99(17):7059-68
  11. Silva AC, Yami M, Libeau G, Carrondo MJ, Alves PM. 2014. Testing a new formulation for Peste des Petits Ruminants vaccine in Ethiopia. Vaccine 32, (24), 2878-2881
  12. Patrone M, Carinhas N, Sousa MQ, Peixoto C, Ciferri C, Carfì A, Alves PM. 2014. Enhanced expression of full-length human cytomegalovirus fusion protein in non-swelling baculovirus-infected cells with a minimal fed-batch strategy. PloS One, 9(3): e90753 
  13. Fernandes F, Dias MM, Vidigal J, Sousa MF, Patrone M, Teixeira AP, Alves PM. 2014. Production of rotavirus core-like particles in Sf9 cells using recombinase-mediated cassette exchange. J Biotechnol, 171 (10): 34–38
  14. Fernandes F, Teixeira AP, Carinhas N, Carrondo MJT, Alves PM. 2013. Insect cells as production platform of complex VLPs. Expert Rev Vaccines. Feb;12(2):225-36.  
  15. Marcelino I, Almeida AM, Brito C, Meyer D, Barreto M, Sheikboudou C, Franco CF, Martinez D, Lefrançois T, Vachiéry N, Carrondo MJ, Coelho A, Alves PM, 2012. Proteomic analyses of Ehrlichia ruminantium highlight differential expression of MAP1-family proteins. Vet Microbiol, 156 (2012) 305–314. 
  16. Rodrigues AF, Guerreiro MR, Santiago VM, Dalba C, Klatzmann D, Alves PM, Carrondo MJ, Coroadinha AS, 2011. Down-regulation of CD81 tetraspanin in human cells producing retroviral-based particles: tailoring vector composition. Biotechnol Bioeng, 108(11):2623-33. 
  17. Silva AC, Carrondo MJ, and Alves PM, 2011. Strategies for improved stability of Peste des Petits Ruminants vaccine. Vaccine, 29:4983-91. 
  18. Roldão A, Mellado MC, Castilho LR, Carrondo MJ, Alves PM, 2010. Virus-like particles in vaccine development. Expert Rev Vaccines, 9(10):1149-76. 
  19. Matias AA, Serra AT, Silva AC, Perdigão R, Ferreira TB, Marcelino I, Silva S, Coelho AV, Alves PM, Duarte CM, 2010. Portuguese winemaking residues as a potential source of natural anti-adenoviral agents. Int. J Food Sci Nutr, 61(4):357-68. 
  20. Adakal H, Stachurski F, Konkobo M, Zoungrana S, Meyer DF, Pinarello V, Aprelon R, Marcelino I, Alves PM, Martinez D, Lefrancois T, Vachiéry N, 2010. Efficiency of inactivated vaccines against heartwater in Burkina Faso: Impact of Ehrlichia ruminantium genetic diversity. Vaccine, 28(29):4573-80. 
  21. Mellado MC, Mena JA, Lopes A, Ramirez OT, Carrondo MJ, Palomares LA, Alves PM, 2009. Impact of physicochemical parameters on in vitro assembly and disassembly kinetics of recombinant triple-layered rotavirus-like particles. Biotechnol Bioeng, 104(4):674-86. 
  22. Ferreira TB, Perdigão R, Silva AC, Zhang C, Aunins JG, Carrondo MJ, Alves PM, 2009. 293 cell cycle synchronisation in adenovirus vector production. Biotechnol Prog, 25(1):235-43. 
  23. Mellado MC, Peixoto C, Cruz PE, Carrondo MJ, Alves PM, 2008. Purification of recombinant rotavirus VP7 for the study of in vitro rotavirus-like particles assembly. J Chromatogr B, 874:89-94. 
  24. Roldão A, Carrondo MJ, Alves PM, Oliveira, R, 2008. Stochastic simulation of protein expression in the Baculovirus/Insect cell system. Comp Chem Eng, 32:68-77. 
  25. Silva AC, Delgado I, Sousa MF, Carrondo MJ, Alves PM, 2008. Scalable culture systems using different cell lines for the Production of Peste des Petits Ruminants Vaccine. Vaccine, 26:3305-11. 
  26. Mellado MC, Franco C, Coelho AV, AlvesPM, Simplício AL, 2008. Sodium dodecyl sulfate-capillary gel electrophoresis analysis of rotavirus-like particles. J Chromatogr A, 1192:166-72. 
  27. Vicente T, Sousa MF, Peixoto C, Mota JP, Alves PM, Carrondo MJ, 2008. Anion exchange membrane chromatography for purification of rotavirus-like particles. J Membr Sci, 311:270-83. 
  28. Ferreira TB, Carrondo MJ, Alves PM, 2007. Effect of ammonia production on intracellular pH consequent effect on adenovirus vector production. J Biotechnol, 129(3):433-38. 
  29. Peixoto C, Sousa MF, Silva AC, Carrondo MJ, Alves PM, 2007. Downstream processing of triple-layered rotavirus like particles. J Biotechnol, 127:452-61 
  30. Peixoto C, Marcelino I, Amaral AI, Carrondo MJ, Alves PM, 2007. Purification by membrane technology of an intracellular Ehrlichia ruminantium candidate vaccine against heartwater. Proc Biochem, 42(7):1084-89. 
  31. Roldão A, Vieira HL, Charpilienne A, Poncet D, Roy P, Carrondo MJ, Alves PM, Oliveira R, 2007. Modeling rotavirus-like particles production in a baculovirus expression vector system: infection kinetics, baculovirus DNA replication, mRNA synthesis and protein production. J Biotechnol, 128(4):875-94. 
  32. Faburay B, Geysen D, Ceesay A, Marcelino I, Alves PM, Taoufik A, Postigo M, Bell-Sakyi L, Jongejan F, 2007. Immunisation of sheep against heartwater in The Gambia using inactivated and attenuated Ehrlichia ruminantium vaccines. Vaccine, 25(46):7939-47. 
  33. Marcelino I, Vachiéry N, Amaral AI, Roldão A, Lefrançois T, Carrondo MJ, Alves PM, Martinez D, 2007. Effect of the purification process and the storage conditions on the efficacy of an inactivated vaccine against heartwater. Vaccine, 25(26):4903-13. 
  34. Lameiro MH, Malpique R, Silva AC, Alves PM, Melo E, 2006. Encapsulation of adenoviral vectors into chitosan-bile salt microparticles for mucosal vaccination. J Biotechnol, 126:152-62. 
  35. Marcelino I, Sousa MF, Veríssimo C, Cunha A, Carrondo MJ, Alves PM, 2006. Process development for the mass production of Ehrlichia ruminantium. Vaccine, 24(10):1716-25. 
  36. Roldão A, Vieira HL, Alves PM, Oliveira R, Carrondo MJ, 2006. Intracellular dynamics in rotavirus-like particles production: Evaluation of multigene and monocistronic infection strategies. Proc Biochem, 41:2188-99. 
  37. Peixoto C, Ferreira TB, Carrondo MJ, Cruz PE, Alves PM, 2006. Purification of adenoviral vectors using expanded bed chromatography. J Virol Methods, 132:121-6. 
  38. Zhang C, Ferreira TB, Cruz PE, Alves PM, Haury M, Carrondo MJ, 2006. The importance of 293 cell cycle phase on adenovirus vector production, Enzyme Microb Technol. 39:1328-32. 
  39. Antonis AF, Bruschke CJ, Rueda P, Maranga L, Casal JI, Vela C, Hilgers LA, Belt PB, Weerdmeester K, Carrondo MJ, Langeveld JP, 2006. A novel recombinant virus-like particle vaccine for prevention of porcine parvovirus-induced reproductive failure. Vaccine, 24(26):5481-90. 
  40. Ferreira TB, Ferreira AL, Carrondo MJ, Alves PM, 2005. Effect of refeed strategies and non-ammoniagenic medium on adenovirus production at high cell densities. J Biotechnol, 119:272-80. 
  41. Vieira HL, Estevão C, Roldão A, Peixoto CC, Sousa MF, Cruz PE, Carrondo MJ, Alves PM, 2005. Triple layered rotavirus VLP assembly: kinetics of vector replication, mRNA stability and recombinant protein production. J Biotechnol, 120(1):72-82. 
  42. Ferreira TB, Alves PM, Aunins JG, Carrondo MJ, 2005. Use of adenoviral vectors as veterinary vaccines. Gene Ther, 12(S1):73-83. 
  43. Ferreira TB, Ferreira AL, Carrondo MJ, Alves PM, 2005. Two different serum-free media and osmolality effect upon human 293 cell growth and adenovirus production. Biotechnol Lett, 27(22): 1809-13. 
  44. Peixoto CC, Marcelino I, Vachiéry N, Bensaid A, Martinez D, Carrondo MJ, Alves PM, 2005. Quantification of Ehrlichia ruminantium by real time PCR. Vet Microbiol, 107:273-8. 
  45. Marcelino I, Veríssimo C, Sousa MF, Carrondo MJ, Alves PM, 2005. Characterization of Ehrlichia ruminantium replication and release kinetics in endothelial cell cultures. Vet Microbiol, 110:87-96. 
  46. Maranga L, Brazão TF, Carrondo MJ, 2003. Virus like particles production at low multiplicities of infection with the baculovirus insect cell system. Biotechnol  Bioeng, 84:245-53. 
  47. Maranga L, Mendonca RZ, Bengala A, Peixoto CC, Moraes RH, Pereira CA, Carrondo MJ, 2003. Enhancement of Sf-9 cell growth and longevity through supplementation of culture medium with hemolymph. Biotechnol Prog, 19:58-63. 
  48. Cruz PE, L Maranga, MJ Carrondo, 2002. Integrated process optimization: lessons from retrovirus and virus-like particle production. J Biotechnol, 99:199-214. 
  49. Maranga L, Rueda P, Antonis AF, Vela C, Langeveld JP, Casal JI, Carrondo MJ, 2002. Large scale production and downstream processing of a recombinant porcine parvovirus vaccine. Appl Microbiol Biotechnol, 59:45-50. 
  50. Maranga L, PE Cruz, JG Aunins, MJ Carrondo, 2002. Production of core and virus-like particles with baculovirus infected insect cells. Adv Biochem Eng Biotechnol, 74:183-206. 
  51. Cruz PE, Peixoto CC, Devos K, Moreira JL, Saman E, Carrondo MJ, 2000. Characterization and downstream processing of HIV-1 core and virus-like-particles produced in serum free medium. Enzyme Microb Technol, 26:61-70. 
  52. Cruz PE, Martins PC, Alves PM, Peixoto CC, Santos H, Moreira JL, Carrondo MJ, 1999. Proteolytic activity in infected and non infected insect cells: degradation of HIV-1 Pr55gag particles. Biotechnol Bioeng, 65:133-43. 
  53. Cruz PE, Peixoto CC, Moreira JL, Carrondo MJ, 1998. Production and quality analysis of Pr55gag particles produced in baculovirus-infected insect cells. J Chem Technol Biotechnol, 72:149-58. 
  54. Cruz PE, Peixoto CC, Moreira JL, Carrondo MJ, 1998. Optimization of the production of virus-like particles in insect cells. Biotechnol Bioeng, 60:408-18.