Monoclonal Antibody Production

Biology

Monoclonal Antibody Production

Monoclonal antibodies (mAbs) are derived from a singe cell. mAbs produce identical offspring and can be reproduced indefinitely.  These antibodies are very specific but can be more difficult and time consuming compared to the production of polyclonal antibodies.

 

Our services are outlined below.

 

 

Option A: Custom monoclonal antibody production using protein antigen

 

  1. Protein Production (optional): We produce at least 2mg of purified protein for immunisation. The customer can also provide their own protein provided we are happy with the purity and buffer components.
  2. Immunisation: We use a standard 46-day immunisation protocol consisting of five immunisations of five mice.
  3. Fusion: Spleen cells are fused with mouse myeloma cells to produce hybridoma cells.
  4. Testing: We send the customer a selection of positively testing clones for them to test under their conditions. The customer will select any that work for them and 2ml of cells and 25ml of cell supernatant will be sent for each clone.

 

 

Option B: Custom monoclonal antibody production using peptide antigen

 

  1. Peptide Design: We select peptides based on the primary and secondary structure of the native protein, and the cross-reactivity with other proteins within the same species.
  2. Peptide Synthesis: Our peptides for immunisation are always >80% pure. We synthesise 18mg in total of the chosen peptide, 2mg of this is aliquoted separately for the customer to use as they wish.
  3. Conjugation to Carrier Proteins: We conjugate the peptide(s) to KLH and BSA for immunisation and testing respectively.
  4. Immunisation: We use a standard 46-day immunisation protocol consisting of five immunisations of five mice.
  5. Fusion: Spleen cells are fused with mouse myeloma cells to produce hybridoma cells.
  6. Testing: We send the customer a selection of positively testing clones for them to test under their conditions. The customer will select any that work for them and 2ml of cells and 25ml of cell supernatant will be sent for each clone.

 

Option C: Custom polyclonal antibody production using a PTM-specific peptide antigen

 

  1. Peptide Design: We select the best possible peptide sequence to include the region of the desired PTM.
  2. Peptide Synthesis: Our peptides for immunisation are always >80% pure.We synthesise 18mg in total of the chosen peptide, 2mg of this is aliquoted separately for the customer to use as they wish. We will also synthesise the required peptide without the PTM in place in order to deplete non-specific antibodies at the purification step.
  3. Conjugation to Carrier Proteins: We conjugate the peptide(s) to KLH and BSA for immunisation and testing respectively.
  4. Immunisation: We use a standard 46-day immunisation protocol consisting of five immunisations of five mice.
  5. Fusion: Spleen cells are fused with mouse myeloma cells to produce hybridoma cells.
  6. Testing: We send the customer a selection of positively testing clones for them to test under their conditions. The customer will select any that work for them and 2ml of cells and 25ml of cell supernatant will be sent for each clone.

 

Interested in any of our services?
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