Custom hybridoma projects are divided into 3 phases each of which can be customized to meet specific client needs. The client has the option of terminating the project prior to the beginning of the next phase.
Animals routinely used for hybridoma development include mice, rats and Armenian hamsters.
All studies are carried out under approved IACUC protocols, USDA Registration, PHS Assurance and AAALAC accreditation.
Immunization of 5 animals including pre-injection bleeds, antigen/adjuvant injections, all antigen/adjuvant boosts, bleeds and ELISA or FACS to determine the animal with the highest titer to be used for cell fusion.
Fusion of spleen cells with myeloma cell line, plating the fusion product into 96 well plates, and determination of those wells expressing antibodies to the antigen by ELISA or FACS assay. Antibody-secreting colonies (up to 48) are isolated and transferred to 24 well plates, expanded and a portion frozen (2 vials). Antibody containing medium (~1ml) of up to 48positive preclones* will be shipped to the client for evaluation in the client’s specific application.
Cloning of antibody-positive wells. A maximum of 5* of the positives selected by the client will be cloned by cell sorting and isotyped. Clones will be expanded, and vials of each selected clone will be frozen. Unpurified antibody rich medium (10-15 ml) from each clone as well as the frozen vials of the cells will be shipped to the client.
* Additional preclones and/or clones can be provided for an additional fee.
Affinity purified antibody from milligram to gram quantities can be produced by our bioreactor facility.
Biacore analysis of antibody/antigen interactions (ka, kd and KD) is also available.