Icosagen|Tokyo Future Style is a bio-tech trading company

HybriFree Technology

The proprietary HybriFree method is used for the discovery of monoclonal antibodies from rabbits and chickens. This includes the capture of antibody-producing cells, cloning of VH/VL sequences and construction of human IgG minilibraries for screening. The antibody expression vectors constructed during antibody discovery can be directly used for the manufacturing in Chinese hamster ovary (CHO) cells.

Benefits of the HybriFree Technology

• Immunization of wild type rabbits and chickens provides great diversity with broad epitope coverage, and picomolar affinities without affinity maturation
• HybriFree in vivo B cell approach captures antibodies with optimal biochemical properties leading to good developability
• Good manufacturability of the antibodies is achieved by using CHO cells for the expression of IgG minilibraries that are developed directly from B cells
• Greater evolutionary distance of chicken yields greater immunogenicity against conserved targets compared to mice
• HybriFree antibody discovery platform is further powered by Icosagen’s patented QMCF expression technology and extensive biochemical competence resulting in high quality antigen production
• Icosagen has a proprietary VLP-technology for high level expression of membrane proteins including GPCRs

Benefits of choosing rabbit or chicken for immunization

• 

  Mouse

  • Slow immunization
  • Normal affinity
  • Industry standard

• 

  Chicken

  • Fastest immunization
  • High affinity
  • The best for conserved targets

• 

  Rabbit

  • Fast immunization
  • Highest affinity
  • The best for small molecules

Benefits to the clients

• Recombinant protein production (up to 50 L scale) is managed by PhD-level scientists that guarantee smooth flow of projects (including difficult to express proteins) and transparent communication with clients, supported by detailed project reports
• Cloning of DNA expression vectors are done with rapid one-step ligase independent method, which is based on Gibson assembly
• Patented QMCF expression technology in CHO and HEK293 cells provides cost-effectiveness and scalability
• QMCF CHO cell pools and production cell banks for the commercial supply of diagnostic antibodies can be generated in 10 days
• Wide range methods for QC analysis are available, including protein concentration, Western blot, PAGE, LabChip, IEF (to determine isoelectric point), analytical SEC (to determine aggregation level), LAL test (to determine endotoxin levels), Octet system (to determine affinity and kinetics), FLOW analysis, DLS (to determine particle size), HILIC HPLC (to determine afucosylation level), LC-MS and LC-MS/MS (to determine proteomic properties)
• Capabilities to develop methods for the purification of native proteins without tags

Icosagen’s protein production service is based on the QMCF technology that is a robust mammalian expression system for recombinant proteins, delivering up to 1.5 g of antibody per litre of media. Expression vectors are constructed and transfected into CHO cells with our proprietary transfection reagent 007. Proteins are purified from the media with the method that is best suited for the protein of interest. Produced proteins and antibodies have high grade of purity, low endotoxin level and native folding. Deliverables include detailed reports.