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.