Multiplexing Phenotype and Function for More Biologically Relevant Insights - 9

High Throughput Combinatorial Profiling of Checkpoint
Inhibitor Antibodies on the Intellicyt® iQue Screener PLUS
Zhaoping Liu and Tom Duensing

C

heckpoint inhibitors have become valuable immunomodulatory targets in the advancement of cancer
treatment. Looking for the synergy between new
checkpoint inhibitor antibodies and known inhibitors is an
important aspect of this research. The iQue Screener PLUS
platform is a powerful tool to simultaneously assess these
interactions in a single well of a microtiter plate. ForeCyt®
software provides plate-level analytics and high content
visualization to generate deep insight rapidly. Using a mixed
lymphocyte reaction (MLR) model, we profiled potential
synergies of several known checkpoint inhibitors antibodies.
Responses of PD-1, PD-L1, and CD73 inhibitors both individually, and in combination with CTLA-4 inhibitors, were
assessed for proliferation, viability and cytokine secretion
simultaneously in the same well. Synergies ranging from 2-10
fold increase over CTLA-4 alone were observed in the secretion of TNF-a and IL-1β. Results were obtained and analysis
completed in a 384-well plate in 30 minutes.
Eight combinations of donor PBMCs were mixed together and
distributed in 384-well plates. The cells were treated for varying
combinations and concentrations of checkpoint modulating
antibodies (anti-PD1, anti-PD-L1, anti-CTLA4, and anti-CD73)
and MultiCyt Cell Proliferation Dye was added. After incubation
for 3 days, reagents for measuring cell viability (MultiCyt Cell
Membrane Integrity Dye) and the secretion levels of 3 different
cytokines (MultiCyt QBeads®) were added to the wells of the
plate. Plates were sampled on the Intellicyt® iQue Screener
PLUS and data was analyzed with ForeCyt Software
9

| January, 2019

Figure 1. Rapid Testing of Anti-CTLA-4 Antibodies in Mixed Lymphocyte Reactions with Multiple Donor Pairs Eight
pairs of donor PBMCs from different donors were treated with increasing concentrations of antibodies specific for
CTLA-4 in triplicate. Data from control compounds, also included on the plate, are not shown. The levels of secreted
cytokines IFNγ, TNFα, IL-1β and cell viability and proliferation were assessed.



Multiplexing Phenotype and Function for More Biologically Relevant Insights

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