Optimize, Adapt and Overcome - 26

Optimize, Adapt, and Overcome: Tips from the Cell Line Development Front * Enhanced Development of Virus-Specific Hybridomas Using ClonePix and CloneSelect Imager Technologies

Figure 4. CloneSelect Imager Software analysis enables identification of optimally
growing clones. Image analysis of the subset of 480 picks demonstrates assessment of
colonies' cellular morphology and growth characteristics (A) as well as estimation of colony
number and colony area for each well (B), thereby efficiently tracking colony growth. Slower
growing colonies were omitted, while the top growers-30% of the total population (146
colonies)-were used to screen for specificity with peptide assay.

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Four hundred and eighty high secretors were picked
from source plates with semi-solid CloneMedia and
deposited into five 96-well destination plates containing 200 µL of liquid cloning media, then allowed to
expand for three days. CloneSelect Imager was used
to image picked clones in 96-well plates in order to
objectively and quantitatively assess cell growth.
Superior imaging speed (less than 90 seconds for
96-well plate) enabled to rapidly monitor and
evaluate colony growth for five days. By effectively
utilizing CloneSelect Imager process, 146 top
growers were isolated (Figure 4).

Screening and Identification of
Sub-Clones for Pilot Scale Analysis
One hundred forty-six top growers were screened for
antigen specificity with a synthesized peptide representing the epitope for mAb binding. The peptide had
exhibited a strong affinity for antibody as demonstrated by previous studies and thereby was used to screen
novel sub- clones secreting high affinity antibodies.

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Optimize, Adapt and Overcome

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