Hybridoma & CHO Clone Picking
A fundamental step in the development of new cell lines for antibody production is the identification of single cell-derived clones that produce high and consistent levels of the target protein. Compared to the time-consuming and labor-intensive steps of conventional techniques, productivity screening with the CellCelector can be performed in a more efficient way.
The antibody generation of hybridoma clones can vary dramatically between different clones. The proteins produced by the clones and secreted into the medium can be made visible by fluorescence-labelled secondary antibodies. As the viscosity of the semi-solid media prevents the antibody from diffusing further into the media the signal will appear as a fluorescent halo around the cell clone that produced it. To select specifically high-producer clones the CellCelector software compares the size of the cell colony visible in bright field with the size of fluorescence halo surrounding the cell colony and calculates the rate of the antibody productivity of each clone. This allows ranking of cell clones across multiple source plates for best antibody production.
Schematic view of hybridoma clones (light blue) and the detection of secreted antibodies (dark blue) by fluorochrome labelled secondary antibodies (green with yellow circle) (A-C)
Microscopic view (overlay of bright field and fluorescence illumination) of hybridoma clones after incubation with fluorochrome labelled secondary antibodies (D-F).
The intensity of antibody production correlates with the diameter of the fluorescent halo around the colony: colony producing no antibodies (A and D), colony producing medium amounts of antibodies (B and E); colony producing high amounts of antibodies equals high producer (C and F). The size of the colonies is similar (see enlarged area in bright field observation D-F).
Antibody producing CHO cell colony; QC images before and after picking with the semi-solid media module of the CellCelector.
CHO cell colony harvested with the semi-solid media module cultured in a 96-well destination plate (at day 1 and day 5 after harvesting).
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