Hybridoma Antibody Development
Phase I: Antigen Preparation and Immunization
• Antigen Preparation/Peptide Conjugation.
• A maximum group of 5 mice will be immunized over a period of 10 weeks. Mouse strain used is BALB/c.
• Serum is screened by ELISA.
• Highest titer screen will be selected for phase II.
Estimated completion time: 12 weeks.
Phase II: Fusion and Selection
• Fusion of splenocytes to myeloma cell line.
• All splenocytes will be used for fusion process.
• Incubation in HAT (Hypoxanthine-Aminopterin-Thymidine) selection media plated in 96 wells up to 10 plates per fusion.
• ELISA to confirm positive antibody.
Estimated completion time: 2 weeks.
Phase III: Screening/Expansion/Cryopreservation
• Antibody Characterization to confirm Monoclonal Antibody.
• Isotype confirmation.
• Cell Culture expansion up to the 10 best clones specific to antigen.
• 3 vials per clone will be frozen and stored in LN2 (Liquid Nitrogen) until shipment.
Estimated completion time: 8 weeks.