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High-Content Screening Assay for the Identification of Antibody-Dependent Cellular Cytotoxicity Modifying Compounds
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JoVE Journal Cancer Research
High-Content Screening Assay for the Identification of Antibody-Dependent Cellular Cytotoxicity Modifying Compounds

High-Content Screening Assay for the Identification of Antibody-Dependent Cellular Cytotoxicity Modifying Compounds

DOI:
10.3791/64485-v

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13:59 min

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August 18, 2023

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, , , , ,
1Department of Medical Chemistry, Faculty of Medicine,University of Debrecen, 2Doctoral School of Molecular Medicine,University of Debrecen, 3National Academy of Scientist Education,University of Debrecen, 4ELKH-DE Cell Biology and Signaling Research Group

Chapters

  • 00:05Introduction
  • 01:02Protocol
  • 01:25Coating of the HCS plate
  • 02:04Seeding of JIMT-1 EGFP cells
  • 04:33Pre-treatment of JIMT-1 EGFP cells with compound library
  • 06:06Starting the ADCC assay by adding the effector cells
  • 09:16Imaging
  • 10:45Image analysis
  • 12:24Representative results
  • 13:15Discussion

Summary

Automatic Translation

Automatic Translation

This protocol presents an automated, image-based high-throughput technique to identify compounds modulating natural killer cell-mediated breast cancer cell killing in the presence of a therapeutic anti-HER-2 antibody.

Tags

Keywords: ADCCAntibody-dependent Cellular CytotoxicityNK CellsGranulocytesMacrophagesEffector CellsFcγR ReceptorsTrastuzumabHER2JIMT-1 CellsNK-92 CellsHigh-content Screening96-well PlateCell CultureCell AttachmentCell CountingTrypan BlueCell SuspensionHigh-throughput ScreeningRobotic ArmPin ToolTest Compounds

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