Multiplexing Phenotype and Function for More Biologically Relevant Insights - 8

ReMore Biologically Relevant Insights | MORE DATA... MORE INSIGHT

we use the Intellicyt® iQue Screener to assess multiple readouts from small sample
volumes, such as cell health and cytokine release in tandem," discusses Rachel Forfar,
Ph.D., senior research scientist, cellular pharmacology, LifeArc.
"The technology is advantageous in its ease of use, the amount of data that can be
generated with little hands-on requirement, and the range of applications that can
be studied. There is no need for prior flow cytometry experience as there are no voltages to set to find your cells or knowledge of which filters to activate. The ForeCyt
software is versatile yet simple to pick up, and the software updates address any
issues raised by customers or added functionality requests."
LifeArc has integrated the use of the IncuCyte and the Intellicyt® iQue Screener
in their Antibody Drug Conjugate programs, from the screening stage to identify
hybridoma supernatants that bind and internalize, to the candidate antibody stage
where full characterization of the binding, internalization and cytotoxicity of toxinconjugated antibodies is undertaken.
Dr. O'Rorke explains that the Intellicyt® iQue Screener is a locked down system.
Due to the wide dynamic range, PMT (photomultiplier tube) adjustments are not
required. This value-add means the screener runs the same every time it is used,
ensuring reproducible data. Color compensation is not required for optimized
reagents offered by Sartorius, adds Dr. ORourke, although it is initially required
for end-user customized assays. After development, an assay and compensation
template can be used to eliminate this requirement.
Software is optimized with standard templates and metrics making the screener plugand-play for routine small partial plate assays or large-scale multi-plate experiments.
"Ease of maintenance and user-serviceable parts has made it possible for our very
small core to keep a high-performance system operating without a lot of extra or
experienced staff. As core lead I have over a decade of experience; however, my two
technicians have little experience and both of them have been able to grasp the software and the maintenance on the instrument with minimal training," points out Ryan.
"Maintenance that might require a dedicated technician or a senior staffer can be
done with well-made wizards and photo instructions provided in the software, making
the Intellicyt® iQue Screener PLUS one of the friendliest machines I have worked with."

| January, 2019

Analyzing Adherent Cells
Kite Pharma desired a HTFC multi-parameter assay for rapid, simultaneous evaluation
of CAR-T cells against adherent cell lines for potential application for the evaluation of
new cell-engineering approaches in the adoptive cell therapy field. Adherent cells are
difficult to analyze on a flow cytometer, which requires a single cell suspension.
To develop the assay, a protocol was needed for getting the adherent cells into
suspension. Multiple assay parameters, including disassociation buffers, incubation
times, shaking methods and times, effector to target ratios, intraplate consistency
of target cell acquisition and function were analyzed and optimized. Results of the
optimized flow cytometric method to characterize transduction efficiency, activation
and cytotoxicity were reported in SLAS Discovery (Figure 2, previous page).3
According to Kite Pharma the Intellicyt® iQue Screener PLUS and assay are also
amenable to further T-cell characterization such as immunophenotyping. The addition of two markers, such as CD45RA and CD62L, could allow users to measure the
composition of naïve central memory, effector memory, and effector T cells. n
References
1. Comparing Flow Cytometry QBeads PlexScreen Assays with Other Immunoassays for
Determining Multiple Analytes. Mei Ding, Anders Cavallin, Nils-Olov Hermansson, Pia
Berntsson, Lisa Jinton, and Sandra Rodrigo Blomqvist. SLAS Discovery, 2018, Vol. 23(7) 676
-686, DOI: 10.1177/2472555218771610
3. Automated High-Throughput Flow Cytometry for High-Content Screening in Antibody
Development. Yana Wang, Tomoki Yoshihara, Samson King, Tinh Le, Patrick Leroy, Xiansi Zhao,
Ching Kit Chan, Zhong-Hua Yan, and Saurabh Menon, SLAS Discovery 2018, Vol. 23(7) 656
-666, DOI: 10.1177/2472555218776607
4. High-Throughput Flow Cytometric Method for the Simultaneous Measurement of
CAR-T Cell Characterization and Cytotoxicity against Solid Tumor Cell Lines. Emily M.
Martinez, Samuel D. Klebanoff, Stephanie Secrest, Gabrielle Romain, Samuel T. Haile,
Peter C. R. Emtage, and Amy E. Gilbert. SLAS Discovery 2018, Vol. 23(7) 603 -612, DOI:
10.1177/2472555218768745

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Multiplexing Phenotype and Function for More Biologically Relevant Insights

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