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Custom Monoclonal Antibody Production

Although BioServ UK treats each contract as a custom project, most of the following elements are applicable and help to ensure a quality product is produced:

  • Consultation with client to discuss antigen selection.
  • Critical assessment of suitable screening methods and relevance to intended end use assay.
  • Ongoing liaison between client and BioServ UK during hybridoma development process.
  • Full confidentiality assured.
  • Project segmentation into different phases to provide researcher flexibility for terminating project at the end of each phase.
     

Phase I: Antigen Design and Immunisation (6–10 wks)

  • Design and preparation of protein or cellular antigens or advice with peptide synthesis and conjugation.
  • Immunisation schedule using BioServ UK SOPs.
  • Determination and quantification of serum titer in end use assay e. g. ELISA, Western Blot, IP.
     

Phase II: Cell Fusion and Screening of Candidate Colonies (3–6 wks)

  • Hybridoma fusion using BioServ UK SOP into 10x96 well plates.
  • Initial screen by ELISA to select strongest positive candidate 'binders'. There is opportunity at this stage to select isotype specific antibodies using defined secondary reagents.
  • Expansion into 24-well plates for harvest of tissue culture supernatant for further antibody screening (ELISA, Western Blot or FACS).
  • Option for client to assess antibody performance: dispatch of 5ml of supernantant from best 10-15 colonies.
     

Phase III: Limiting Dilution Cloning and Cryogenic Preservation (3–5 wks)

  • Limiting dilution cloning performed with strongest parental colonies.
  • Subclones with high specificity expanded and cryopreserved (5 vials each).
  • Shipment of hybridoma cells and up to 5ml of supernatant from each subcloned cell line.
     

Phase IV: Large Scale Antibody Production (3–6 wks)

  • Milligramme to gramme quantities produced.
  • Production performed in vitro using a variety of bioreactor systems with option for protein free adaptation.
  • Purification using protein A or G affinity chromatography.