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Colony formation

Cells are grown in source plates containing semi-solid CloneMedia. This approach facilitates plating out of large numbers of cells and ensures formation of discrete colonies for subsequent recovery as independent clones.

Colonies of commonly used cell types growing in CloneMedia. Images captured using CloneSelect Imager

screening CHO cells, clonal screening, rapid clonal screening
CHO colonies, serum-free suspension-adapted, in CloneMedia-CHO imaged on day 8 post-plating.
screening hybridoma colonies
Hybridoma colonies in CloneMedia-Hybridoma/Myeloma imaged on day 8 post-plating.
screening HEK 293 colonies
HEK293 colonies, serum-free, suspension-adapted, in CloneMedia-HEK imaged on day 13 post-plating.
screening CHOK1SV colonies
CHOK1SV colonies, serum-free, suspension-adapted, in CloneMedia-CHOK1SV imaged on day 9 post-plating.

Choice of detection methods

Detection using white light for imaging and picking, fluorescence for indicating expression levels.

Detect recombinant proteins and expression markers

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The protein construct contains epitope tag(s) e.g. His, FLAG® or Fc using tag-specific agents.

Label-free detection of antigen-specific mAbs from hybridomas

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Fluorescently-conjugated or tagged antigen plus CIF (complex initiation factor) are used to generate precipitation.

Label-free detection of secreted antibodies

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CloneDetect fluorescently- conjugated agents enable in situ detection of secreted mouse, rat or human IgG. These agents can be added as liquids or simply sprayed onto the colonies. Relative secretion levels are then measured across 1000’s of clones in parallel.

 

Imaging and analysis

Colonies are imaged in white light and fluorescence then analyzed by the system.

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White light imaging identifies and measures clone morphology, size and proximity
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Fluorescence imaging identifies levels of expression

Ranking and data display

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Ranking plot
  • Clones ranked according to fluorescence
  • User defines selection parameters – system selects clones
    • Morphometric criteria such as size, roundness and proximity to neighbors
    • Quantitative criteria such as protein secretion or specific protein production
  • Bar-coded source and destination plates enable data tracking.
  • Images of all picked clones taken before and after picking are stored with their data, including picking coordinates

Accurate picking and transfer

ClonePix selects colonies, transferring each colony into one well of a 96-well destination plate for growth assay and/or expansion of clonal cells.

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Brochures
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ClonePix™ 2
Screening and selection of mammalian clones
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Accelerate Cell Line Development
ClonePix with CloneSelect Imager
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Recorded Webinars

Using high throughput automated selection to increase discovery of rare, high producing mammalian cell lines

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Optimal selection of monoclonal cell lines producing high amounts of biopharmaceutical human IgGs

Dr Gerritsen, Technology Expert, explains how Genmab have optimized and integrated emerging technologies to achieve an optimal, state-of-the-art process for generation of CHO cell lines expressing therapeutic IgGs.
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