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Biacore spr binding analysis
( A ) CF647-labeled ImmTAAI molecules were assessed for pHLA binding and compared to unlabeled molecules by <t>SPR</t> <t>using</t> <t>BIAcore</t> 8K. Binding to PPI or Tel cognate pHLA was carried out at 37°C. ( B ) Similarly, CF647-labeled ImmTAAI molecules were assessed for PD-1 binding and compared to unlabeled molecules. PD-1 binding was carried out at 25°C. ( C ) Schematic of the ECN90 beta cell line: Jurkat NFL Mel5 PD-1 reporter assay. ( D ) ECN90 or NCI-H1703 cells were pulsed with Melan-A–activating peptide, and titrations of unlabeled or labeled PPI ImmTAAI molecules were added.
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( A ) CF647-labeled ImmTAAI molecules were assessed for pHLA binding and compared to unlabeled molecules by SPR using BIAcore 8K. Binding to PPI or Tel cognate pHLA was carried out at 37°C. ( B ) Similarly, CF647-labeled ImmTAAI molecules were assessed for PD-1 binding and compared to unlabeled molecules. PD-1 binding was carried out at 25°C. ( C ) Schematic of the ECN90 beta cell line: Jurkat NFL Mel5 PD-1 reporter assay. ( D ) ECN90 or NCI-H1703 cells were pulsed with Melan-A–activating peptide, and titrations of unlabeled or labeled PPI ImmTAAI molecules were added.

Journal: Science Advances

Article Title: Beta cell–targeted PD-1 agonist inhibits cell-mediated autoimmunity in pancreas tissue slices

doi: 10.1126/sciadv.aec9029

Figure Lengend Snippet: ( A ) CF647-labeled ImmTAAI molecules were assessed for pHLA binding and compared to unlabeled molecules by SPR using BIAcore 8K. Binding to PPI or Tel cognate pHLA was carried out at 37°C. ( B ) Similarly, CF647-labeled ImmTAAI molecules were assessed for PD-1 binding and compared to unlabeled molecules. PD-1 binding was carried out at 25°C. ( C ) Schematic of the ECN90 beta cell line: Jurkat NFL Mel5 PD-1 reporter assay. ( D ) ECN90 or NCI-H1703 cells were pulsed with Melan-A–activating peptide, and titrations of unlabeled or labeled PPI ImmTAAI molecules were added.

Article Snippet: SPR binding analysis was performed using a BIAcore 8 K (GE Healthcare) as described previously ( ).

Techniques: Labeling, Binding Assay, Reporter Assay