Package details

 
  • Antigen design (design of the most appropriate fragment to be produced)
  • Two gene syntheses (each gene optimized for a specific expression system) and two subclonings
  • Expression tests in two systems taken from E. coli, B. subtilis, , insect cells, and mammalian cells. To be determined by ProteoGenix. GFP-tagged version also done in parallel to check Flow Cytometry (FC) staining specificity
  • Production of the antigen in the most appropriate expression system, depending on the yield and the project requirements.
  • Mice injections (5 mice)
  • Immune response control by FC with transfected cells with GFP obtained at antigen expression test step
  • Selection of the best two mice for two fusions, in order to have a large number and variety of clones
  • Screening of the fusion product by ELISA: around 10 x 96-well plates
  • Screening of the positive clones by FC on transfected cells with specificity control via GFP co-staining
  • Subcloning of best 5 parental clones: 2-3 rounds of limiting dilutions and screening by ELISA
  • Screening of best positive clones by FC on transfected cells with specificity control via GFP co-staining
  • Production and purification of 2mg from the best clone and QC by flow cytometry
  • Level 1 : If we don’t succeed in producing an antibody that detects the immunogen in FC, we charge EUR 1000
  • Level 2 : If we succeed in producing an antibody that works in FC against the immunogen, we charge a part of the cost
  • Level 3 : If after testing the purified antibody the customer wants to receive the hybridoma, we charge the full package price
  • 10 -20 best clones at parental stage for customer’s selection
  • 100 µg of purified antibody to confirm hybridoma purchase
  • 50 µg of purified protein
  After total payment:
  • 1.9 mg of purified antibody
  • One hybridoma cell line (four frozen vials)
  • 150 µg of purified protein
  • Gene in pUC57
  • Gene synthesis: 3-4 weeks
  • Protein production : 7-9 weeks
  • Injections: 8-12 weeks
  • Fusion & screening: 5-6 weeks
  • Subcloning: 8-12 weeks
  • Antibody production: 3-4 weeks
  • Total: 34-47 weeks