Hybridoma colonies - Single Cell Cloning Hybridoma

Single Cell Cloning Hybridoma

Cell3iMager duos scanning technology can be effectively used for selection and monitoring of single cell clonal growth into colony formation

Introduction

Stable single clone isolation technique is highly useful for selecting cells secreting the desired antibody for large scale production. Cell 3imager DUOS imaging platform can be utilized for label free assessment of single cells, picking up the clone of interest and further assessment of the desired clone over a period of days. This allows monitoring the growth of single cell to a full fledged colony.

Hybridoma colonies - Single Cell Cloning Hybridoma
Hybridoma 4B2 colony derived from single cell

Material and Method

  • Cell Line: Hybridoma 4B2 cells (ATCC)
  • Medium: RPMI (ATCC)
  • Plate: 96-well plate flat bottom (CORNING)
  • Seeding cell density: 0.78 cells/well
  • Culture days: 7 days
  • Imaging methods:

    • Bright field
    • (Day 0 and 2) High magnification lens
    • (Day 7) Low magnification lens

Result and Conclusion

  • A well (ex. position ‘E-4’) could be confirmed that the colony derived from single cell.
  • Cell3iMager duos is possible to detect between from 'single cell' to 'colony'.
  • Since it is a label-free analysis, the time-dependent change of same samples can be measure.
Single Cell Cloning Hybridoma - Hybridoma colonies
Cell3iMager duos is possible to detect between from 'single cell' to 'colony'.

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