Monoclonal antibodies

Development of monoclonal antibodies

Gemacbio develops personalized monoclonal antibodies at the request of customers. the main requests concern small-sized molecules and peptides. The development of monoclonal antibody is divided into 3 main steps:

• Immunisations and characterization on mouse

- Cross linking of small molecule (or peptide) to proteins carriers
- Immunisation of five Balb/c mice
- Evaluation of antibodies titer, affinity and specificity in mouse serum by ELISA
- Selection of the best mouse to be used for cell fusion

• Cell fusion and screening

- Fusion of the spleen cells of the selected mouse with the myeloma cell line SP2O in order to generate hybridoma primary cultures
- Plating the fusion into six 96-well plates
- Cultivation on selective media
- Screening by ELISA of wells producing specific antibodies against the induced antigen
- Transfer of the positive primary cultures to 24-wells plates for cell propagation
- Confirmation of antibodies production by the selected primary cultures and check for affinity and specificity using competitive ELISA

• Cloning and final characterization

- Cloning of 4 of the best selected primary culture by limited dilution
- ELISA Screening of the cloned cultures and selection of the highest positive clones for recloning
- Recloning,
- Propagation of 2 final clones
- Cryoconservation
- Isotype determination
- Shipping of 3 cryo-stored vials (on dry ice) for each clone
- Production and shipping of 1mg of protein G-purified antibody for each clone.

For all questions about price and design of these products, please send us an email specifying your needs. We will send you back a detailed estimate as soon as possible. ab [at] gemacbio [dot] com