Mouse Monoclonal Antibody Production

Our Mouse Monoclonal Antibody Production package can be tailored to meet your specific project requirements. Our standard hybridoma generation package includes:

  • Antigen production or conjugation (if required)
  • Immunization of 5 mice
  • Cell fusion
  • Screening by Dot Blot and ELISA
  • Supply of positive clones for customer evaluation
  • Propagation of hybridomas
  • Recloning
  • Expansion & supply of at least 2 clones to the target

Each stage of this protocol is fully adaptable to customer requirements to enable us to provide the mouse monoclonal antibody that is most suitable for your experiment.

Mouse Monoclonal Antibody Production Package Details:

 

Antigen preparation
We can generate small peptides to your specifications, or use your pre-prepared antigen (>90% purified, and ready for injection).  If necessary we can conjugate your chosen peptide to a carrier protein, such as CCH, KLH or BSA  to maximise the immune response.

 

Immunization
5 mice are immunized 4 times per project, using roughly 50 µg of antigen each time. This stage of the protocol is adaptable to your requirements, and we will endeavour to accomodate your protocols. Test bleeds are performed shortly after each immunization. We can send these bleeds to you for evaluation, or evaluate them for you.

 

Cell fusion
The best 2-3 immunized animals are selected from the original 5 that were immunized. The splenocytes are harvested and fused with suitable myeloma cells.

 

Screening
Screening is performed by ELISA and BioDOT as standard, using non-specific Ig and the specific antigen. We will also use another cross reactant of your choosing. At the same time we will provide samples of the positive hybridomas to you for your evaluation.

 

Propogation of hybridomas and recloning
Upon completion of screening, cryopreservation and production of master cell banks takes place. All the positive clones are recloned, and Ig characterization (IgG1, IgG2 etc.) occurs.

 

Expansion
We produce 50 ml of supernatant from each clone and send it to you for final testing. We will also prepare further cell banks from each cloned hybridoma.


Additional Mouse Antibody Services

Upon generation of your polyclonal hybridoma, we can offer additional services to meet your individual requirements. The cost of each service is dependent on the scope of the project.