Development of a Bioluminescent Cell-Based Assay to ... - Promega
Development of a Bioluminescent Cell-Based Assay to ... - Promega
Development of a Bioluminescent Cell-Based Assay to ... - Promega
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Signaling pathways activated in effec<strong>to</strong>r cells when<br />
FcgRIIIa is bound by target cell-bound antibody in ADCC<br />
FcgRIII initiated signaling events<br />
Leibson-PJ, Immunity 1997<br />
Confidential and Proprietary. Not for Further Disclosure.<br />
New reporter-based bioassay<br />
New reporter gene bioassay uses<br />
antigen<br />
FcgRIIIa V158<br />
an earlier step in the pathway<br />
Target cell<br />
Read out: NFAT-RE reporter<br />
• Avoids preparation and high<br />
activation from effec<strong>to</strong>r cells<br />
variability <strong>of</strong> primary effec<strong>to</strong>r cells<br />
ADCC cy<strong>to</strong><strong>to</strong>xicity bioassay<br />
• Avoids high background and<br />
FcgRIIIa<br />
variability <strong>of</strong> spontaneous lysis<br />
Target cell<br />
Antibody<br />
Engineered<br />
Jurkat cell<br />
• To quantify biological activity <strong>of</strong><br />
Antibody<br />
Abs in this MOA pathway = NFAT-RE-luc<br />
Primary<br />
NK cell<br />
Traditional readout is cy<strong>to</strong><strong>to</strong>xicity,<br />
the Read ultimate out: target endpoint cell cy<strong>to</strong><strong>to</strong>xicity<br />
3
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