Our Smart Selection Approach
Hybridomas are immortal somatic cell hybrids that secrete antibodies. Hybridoma generation is a critical step in making monoclonal antibodies. A major barrier to monoclonal antibody discovery is hybridoma selection. Hundreds to thousands of hybridoma cells have to be manually screened in the hope of identifying antibody-secreting clones.
BiCell Scientific Inc has developed a “smart” selection approach to identify antibody-secreting hybridoma cells and streamlined the process for monoclonal antibody production.
Flow chart of “smart” hybridoma selection
Step 1:
Mouse is immunized with antigen
Step 2:
Splenocytes from immunized mouse are isolated
Step 3:
Splenocytes are fused with myeloma cells (our electrofusion protocol can generate 1 hybridoma clone out of 104 splenocytes)
Step 4:
Hybridoma cells are treated by a proprietary crosslinking protocol to immobilize antibodies on the cell surface
Step 5:
Fluorophore labeled antigen molecules allow identifying positive hybridoma cells by direct binding to the hybridoma cell surface
Step 6:
Fluorescence based cell sorting
Step 7:
Dilution cloning and clonal selection
Electrofusion efficiently fuses myeloma cells with splenocytes. Circle indicates fusion event. Myeloma cells: large nuclei; splenocytes: small nuclei.
Antigen labeling of hybridoma cells. Hybridoma cells are transiently crosslinked to immobilize antibodies on the cell surface. FITC-labeled antigen binds to and decorates the hybridoma cell that secretes the correct antibody.