Cloning in Monoclonal Antibody Discovery

Monoclonal antibodies depend on cloning for establishment of monoclonal hybridoma cultures, i.e. cultures obtained from a single antibody-producing hybridoma cell. We use fluorescence-activated cell sorting (FACS) to accurately create single monoclonal cultures. Labs performing cloning by limiting dilution can not guarantee mono-clonality of the cultures and may loose a high percentage of the initial “hits”. Our monoclonal cultures show high recovery of the expanded monoclonal culture (>95%) and do not required multiple rounds of sub-cloning. Additionally, false negative or positive cultures are minimized since only one antibody is present in each culture. We typically clone from 10-30 96-well plates of hybridomas. The cloning and expansion process typically takes 3 weeks.

the-flow-cytometer
    Flow sorting of single viable hybridomas:

  • Ensures monoclonality of each culture tested.
  • Achieves 95% recovery of positive clones
  • Better recovery than limited dilution cloning.

cloning