Steps | Service Content | Cycle (weeks) |
Antigen Preparation | Codon optimization and gene synthesis | 2 |
Protein expression, purification, and scale-up production (4mg) | 2 | |
Animal Immunization (5 mice) | Primary immunization: 4 doses Optional boosters: 1-2 doses Antibody titer monitoring by ELISA Provision of immune sera from 5 immunized mice for client validation | 4 |
Mouse cell fusion and re-screening | Splenocyte isolation and cell fusion using the mouse exhibiting the highest serum antibody titer | 2 |
Subcloning and Screening | Monoclonal hybridoma development protocol: Limiting dilution subcloning (0.5 cells/well) Iterative screening (2-3 cycles) ELISA testing of 96 supernatants Expansion of 10+ stable monoclonal lines Provision of supernatants (1mL/line) for client validation | 2 |
Stable clone establishment, cryopreservation, and antibody production | Selection of one cell line for expansion based on screening results, followed by antibody purification (2mg) and ELISA titer determination | 4 |
Total | 3 months | |
