New clonality reporting for Solentim Cell Metric; facilitates documentation of clones for regulatory reporting process

1. Clonality Assurance
Exportable Documentation using the Cell MetricTM CLD
July 2015

2. Needs
• FDA requirements for image documentation (reference -Sarah Kennett)
• Image of the entire well needed
• Is there 1 cell or 2 cell at Day 0
• Single colony image not sufficient (single cell image needed)
• This proof data is needed as part of IND and BLA for biologicals and
biosimilars
• Current customer methods
• Manual with customers currently cutting, pasting & annotating image data
• No standard formats
• Loss of audit trail
• Takes a long time to put together

3. Clonality Report
• Report function within the Cell Metric CLD software for creation and publication
of a clonality report.
• This report tracks all growth time points for a given well containing vital
information such as a single cell loci, cell division, colony growth, plate debris or
artefacts and other features of interest.
• Free of charge upgrade for existing users

4. The ability to go back in time - Recommended order of
image review
• Day 14 (approx.)
• Scan to identify wells with good growth characteristics (% confluence)
• Day 7
• Intermediate scan to identify number of colony loci growing in a well
• Day 0
• Post seeding image review of only the wells showing desired growth characteristics. Choose
based on external productivity data and viability data combined with confluence data from
day 14 and loci information from day 7. look for single cell in the well
• Pre-scan to capture images of wells before seeding to identify debris
• Day 1
• Confirm single cell at day 0 is now two cells
• Day 2
• Two cells should now be four cells (based on binary divisions of both cells)

5. Features of report page
• Generated from ‘within’ the Cell Metric software, no cutting and
pasting
• Timeline of selected time points as whole well thumbnails
• One time point = one page of report
• Zoomed in on features of interest
• Main Feature is single cell locus
• Fully user-editable naming and annotation of features – cells, debris,
other
• Minutes (rather than days) to compile this report

10. Practical Notes
• Cell Metric CLD has the advantage that 100% of the wells are always
in focus (regardless of whether cells are present)
• Where some of our customers use FACS to dispense cells into culture
media, they do a pre-scan image of plates + media (before cells
added)
• The pre-scan time point can then included in the clonality report
• This is a rapid way to eliminate debris and artefacts as they were
present before the cell was added

11. Practical Notes
• Useful to use the high resolution “verify clonality” imaging mode on
Day 0, 1, 2 to see single cell and then audit initial binary divisions:
- 1 cell becomes 2 cells
- 2 cells become 4 cells
- Odd numbers at this early stage are a warning sign
• Additionally want to see a single colony outgrowth forms from a
single cell progenitor

12. Format of the report
• Output in Powerpoint, PDF or Print
• Can be generated at the PC terminal of the Cell Metric or elsewhere
using remote data viewer software option
• Takes minutes rather than hours or days to generate

13. Existing Pharma customer poster at ESACT
2015 – testimonial (ref)
‘This report function enables the operator an exportable
option which is “proof that can travel” in paper reports,
electronic files and presentation’

14. References
• “Establishing clonal cell lines – a regulatory perspective” Sarah Kennett,
FDA, 2014
• Cell (CHOZN) image data courtesy of SAFC Bioscience
• ESACT 2015 poster # P-1.69 “Implementation of new methods for single
cell sorting and plate imaging results in an improved cell line
development platform for biologics development” Cairns et al. (Sanofi)