I have immunized BalB/C mice with a protein using the intradermal (ID) method with Complete Freund's Adjuvant (CFA) and Incomplete Freund's Adjuvant (IFA), following a 14-day interval and three booster doses for monoclonal antibody development using single B cell cloning. After isolating the plasma cells using MACS, I found that 90% of the plasma cells are producing IgM. However, I need IgG-producing plasma cells.
How can I increase the proportion of IgG-producing plasma cells? Could you provide some suggestions on immunization strategies to achieve a higher yield of IgG-secreting plasma cells?
Thanks for your interesting question, but did you try to use any other method like CHO or Recombinant DNA technology other than Hybridoma technology?
Karuppiah Valliappan
Your antigen may be antigen independent:
If you already tried to optimize the doses, number of boosts and schedule.
You can try to prime with a different adjuvant.
But if your protein is t independent you'll probably need to couple it to a carrier protein to elicit the T response.
Some post immunization intervention can be performed like administration of IL4 or IL12 to influence class switching to IgG, but i can't really say about this one.