A High Throughput, Multiplex Antibody Internalization Assay

Watch the video narration of a poster which describes a high throughput, multiplex, no-wash assay that measures antibody internalization (ABI), specificity and cell health, from a single 30 µL assay volume in 96-and 384-well formats.

Antibodies are easily labeled with the novel, pH-sensitive ABI reagent. The labeled antibodies have little fluorescence at neutral pH, but become highly fluorogenic at low pH when internalized and processed through the acidic lysosome/endosome pathway.

Cell viability is simultaneously measured using cell membrane integrity dye, and cell specificity is characterized using encoding dye. Data acquisition is performed using the iQue^® advanced flow cytometry platform and integrated Forecyt^® software, which combine high throughput sampling, flow cytometry detection and plate level analytics. This delivers rich content with sampling times of less than 20 minutes for a 384-well plate.