title: Cancer vaccine characterization: from bench to clinic.
abstract: BACKGROUND
  The development of safe, effective, and affordable vaccines has become a global effort due to its vast impact on overall world health conditions. A brief overview of vaccine characterization techniques, especially in the area of high-resolution mass spectrometry, is presented. It is highly conceivable that the proper use of advanced technologies such as high-resolution mass spectrometry, along with the appropriate chemical and physical property evaluations, will yield tremendous in-depth scientific understanding for the characterization of vaccines in various stages of vaccine development. This work presents the physicochemical and biological characterization of cancer vaccine Racotumomab/alumina, a murine anti-idiotypic antibody that mimics N-glycolyl-GM3 gangliosides. This antibody has been tested as an anti-idiotypic cancer vaccine, adjuvated in Al(OH)3, in several clinical trials for melanoma, breast, and lung cancer.
  
  
  METHODS
  Racotumomab was obtained from ascites fluid, transferred to fermentation in stirred tank at 10 L and followed to a scale up to 41 L. The mass spectrometry was used for the determination of intact molecule, light and heavy chains masses; amino acids sequence analysis, N- and C-terminal, glycosylation and posttranslational modifications. Also we used the DLS for the size distribution and zeta potential analysis. The biological analyses were performed in mice and chickens.
  
  
  RESULTS
  We observed differences in glycosylation pattern, charge heterogeneity and structural stability between in vivo-produced and bioreactor-obtained Racotumomab products. Interestingly, these modifications had no significant impact on the immune responses elicited in two different animal models.
  
  
  CONCLUSIONS
  We are demonstrated that this approach could potentially be more efficient and effective for supporting vaccine research and development.
authors.affiliation: Process Development Direction, INIM, Center of Molecular Immunology, PO Box 11600, Havana, Cuba. Electronic address: katiar@cim.sld.cu.
authors.forename: K
authors.initials: K
authors.lastname: de la Luz-Hernández
authors.affiliation: Process Development Direction, INIM, Center of Molecular Immunology, PO Box 11600, Havana, Cuba.
authors.forename: Y
authors.initials: Y
authors.lastname: Rabasa
authors.affiliation: Center of Genetic Engineering and Biotechnology, PO Box 16017, Havana, Cuba.
authors.forename: R
authors.initials: R
authors.lastname: Montesinos
authors.affiliation: National center for Laboratory Animal Breeding, Havana, Cuba.
authors.forename: D
authors.initials: D
authors.lastname: Fuentes
authors.affiliation: Process Development Direction, INIM, Center of Molecular Immunology, PO Box 11600, Havana, Cuba.
authors.forename: J F
authors.initials: JF
authors.lastname: Santo-Tomás
authors.affiliation: Process Development Direction, INIM, Center of Molecular Immunology, PO Box 11600, Havana, Cuba.
authors.forename: O
authors.initials: O
authors.lastname: Morales
authors.affiliation: Process Development Direction, INIM, Center of Molecular Immunology, PO Box 11600, Havana, Cuba.
authors.forename: Y
authors.initials: Y
authors.lastname: Aguilar
authors.affiliation: Process Development Direction, INIM, Center of Molecular Immunology, PO Box 11600, Havana, Cuba.
authors.forename: B
authors.initials: B
authors.lastname: Pacheco
authors.affiliation: Process Development Direction, INIM, Center of Molecular Immunology, PO Box 11600, Havana, Cuba.
authors.forename: A
authors.initials: A
authors.lastname: Castillo
mesh_terms: D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000737:chemistry*; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D058846:Antibodies, Monoclonal, Murine-Derived; D019149:Bioreactors; D019496:Cancer Vaccines / Q000737:chemistry*; D002645:Chickens; D002851:Chromatography, High Pressure Liquid; D005285:Fermentation; D006031:Glycosylation; D013058:Mass Spectrometry; D051379:Mice; D010084:Oxidation-Reduction; D010316:Particle Size; D010449:Peptide Mapping; D011499:Protein Processing, Post-Translational; D013678:Technology, Pharmaceutical / Q000379:methods; D064166:Vaccine Potency
chemical_list: D000888:Antibodies, Anti-Idiotypic; D000911:Antibodies, Monoclonal; D058846:Antibodies, Monoclonal, Murine-Derived; D019496:Cancer Vaccines; C000590234:racotumomab
keywords: Cancer vaccine; Comparability studies; Mass spectrometry; Monoclonal antibody