Immunizing Mice for Monoclonal Antibody Production

1. Mix 0.15 mL of 0.5 mg/mL solution of antigen with 0.15 appropriate adjuvant and mix (volumes based on group of three mice).

2. Inject 0.1 mL of adjuvant/antigen mixture per dose intraperitoneally (IP).

4. Obtain test bleeds by tail tip amputation under light anesthesia on d 54 (see Note 5) and assess for antibody titre. Mice should be marked by ear punching or tattooing to allow subsequent identification.

5. Rest the mice for a period of 60 d or more to allow the B-cells to become quiescent.

6. Inject (boost) the best responding mouse with an IP injection of 0.05 mL antigen (0.5 mg/mL solution) without adjuvant.

7. Kill the mouse 3 d later by cervical dislocation and remove its spleen aseptically for cell harvesting.

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