Immuno-Oncology Advances in Applied Cellular Analysis - 22

Advances in Applied Cellular Analysis * Continuous Live-Cell Analysis of the Immune-Tumor Axis

proliferation, and a concomitant rise in apoptotic
tumor cell death, with an IC50 value of 8ng.ml-1.
The three biosimilar mAbs each produced comparable killing effects to trastuzumab, with IC50
values in the range 5-9ng.ml-1. Z' values ranged
from 0.63-0.83 verifying assay robustness. Inspection of the image-and video-sets for each group
verified the immune attack of tumor cells, the
morphological hallmarks of apoptotic cell death
(e.g., degranulation, nuclear condensation), and
the fluorescent signal integrity (target vs. effector
cells). Together, these data provide strong, direct
evidence of biosimilarity for these antibodies for
ADCC in human cells.

CD47-Tumor Cell Engulfment Assay
in Human Bone-Marrow Derived
Macrophages
CD47 is a ubiquitously expressed immuneregulatory protein best known for its "don't eat
me" function that prevents phagocytic removal of
healthy cells. Many cancerous cells express high
levels of CD47, thereby circumventing anticancer
immune responses. Based on this, CD47 has
become a prominent target in the field of cancer
22

| GENengnews.com

Figure 5. Quantification of trastuzumab biosimilar antibodies for antibody-dependent cell killing (ADCC)
using IncuCyte continuous live-cell analysis.
Timecourse and concentration-response curves for tumor cell number (A) and apoptotic target cell death
measured using caspase-3/7 substrate (B). Concentration values in legend are ng.ml-1. Microplate views
for four 96-well plates with derived Z' parameters from the high and low control values (C).

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Immuno-Oncology Advances in Applied Cellular Analysis

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