clonality report for cell metric customer launch

Clonality AssuranceExportable Documentation using the Cell MetricTM CLD

July 2015

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

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

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)

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

Clonality ReportBatch Page

Plate Well Image

Demonstration 1 of 4

CHOZN plated 022315 B2 23/02/2015 21:00

Imagery and report obtained using Solentim Cell Metric Report generated 15/06/2015 12:47

Whole Well

Main Feature

Single cell

Other Features

Debris

Debris

1mm

500µm

50µm

100µm

Well Timeline

23/02/2015 21:00(Hour 2)

24/02/2015 15:12(Hour 20)

02/03/2015 16:41(Day 7)

09/03/2015 15:21(Day 14)


Plate Well Image


CHOZN plated 022315 B2 24/02/2015 15:12


Whole Well

Main Feature

Single cell divided

Other Features

Debris

Debris

1mm

500µm

50µm

100µm

Well Timeline

23/02/2015 21:00(Hour 2)

24/02/2015 15:12(Hour 20)

02/03/2015 16:41(Day 7)

09/03/2015 15:21(Day 14)


Plate Well Image


CHOZN plated 022315 B2 02/03/2015 16:41


Whole Well

Main Feature

Colony from single cell

Other Features

Debris

Debris

1mm

500µm

50µm

100µm

Well Timeline

23/02/2015 21:00(Hour 2)

24/02/2015 15:12(Hour 20)

02/03/2015 16:41(Day 7)

09/03/2015 15:21(Day 14)


Plate Well Image


CHOZN plated 022315 B2 09/03/2015 15:21


Whole Well

Main Feature

Colony

1mm

500µm

Well Timeline

23/02/2015 21:00(Hour 2)

24/02/2015 15:12(Hour 20)

02/03/2015 16:41(Day 7)

09/03/2015 15:21(Day 14)

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

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

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

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’

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)

clonality report for cell metric customer launch

Technology