KILR™ Cytotoxicity Assays and Retroparticles
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The KILRTM cytotoxicity assay specifically measures target cell death in co-cultures. It provides an easy-to-use and sensitive method to determine target cell death with chemiluminescent readout. The assay has broad applications in developing cancer immunotherapies like ADCC, ADCP, CAR-T, and bi-specific antibody therapies. It offers high specificity, sensitivity, flexibility and reproducibility compared to other cytotoxicity assays.
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KILR™ Cytotoxicity Assays and Retroparticles
- 1. 11 KILR™ Cytotoxicity Assays Specifically Measure Target Cell Death in Co-cultures
- 2. 22 Challenge For Developing Cancer Immunotherapies • Specifically determine target cell death in co-cultures with immune effectors? • How to develop assays with multiple target cells? ADCC ADCP CAR-T T-cell Redirection With Bi-Specific Ab Adoptive T-Cell Therapy
- 3. 33 Challenges with Existing Cytotoxicity Assays • Dye-based assays are radioactive and/or leaky • General cell death measures are not specific for target cells • Assays may not reflect the true Mechanism of Action of the drug • Low throughput of assays
- 4. 44 Introducing the KILR™ Cytotoxicity Assay +
- 5. 55 KILR™ Cytotoxicity Assay Highlights & Benefits • Unparalleled Specificity • Signal only from dead target cells • Easy to Use • Simple, add and read protocol with chemiluminescent readout • Exquisite Sensitivity • Detect as few as 75 dead cells/well • Ultimate Flexibility • Run cytotoxicity assays from 30 minutes to 72 hours Benefit Low background and higher signal to noise ratios
- 6. 66 KILR™ Cytotoxicity Assay Highlights & Benefits • Unparalleled Specificity • Signal only from dead target cells • Easy to Use • Simple, add and read protocol with chemiluminescent readout • Exquisite Sensitivity • Detect as few as 75 dead cells/well • Ultimate Flexibility • Run cytotoxicity assays from 30 minutes to 72 hours Benefit Rapid implementation in any lab & eliminate radioactivity
- 7. 77 KILR™ Cytotoxicity Assay Highlights & Benefits • Unparalleled Specificity • Signal only from dead target cells • Easy to Use • Simple, add and read protocol with chemiluminescent readout • Exquisite Sensitivity • Detect as few as 75 dead cells/well • Ultimate Flexibility • Run cytotoxicity assays from 30 minutes to 72 hours ADCC assay with Cetuximab & PBMCs Benefit Low false negatives in a screen and better lead optimization
- 8. 88 KILR™ Cytotoxicity Assay Highlights & Benefits • Unparalleled Specificity • Signal only from dead target cells • Easy to Use • Simple, add and read protocol with chemiluminescent readout • Exquisite Sensitivity • Detect as few as 75 dead cells/well • Ultimate Flexibility • Run cytotoxicity assays from 30 minutes to 72 hours Benefit Develop assay time based on relevant drug biology
- 9. 99 Broad Applications of KILR™ Assays ADCC ADCP CAR-T T-cell Redirection With Bi-Specific Ab Adoptive T-Cell Therapy • Cell death endpoint • Easy-to-use • Reproducible for QC • Cell death endpoint • Plate-based assay • Easy-to-use • Target cell specific • Target cell specific • Run assay for 72 hrs • Easy-to-use • High throughput • Reproducible for QC • Target cell specific • Run assay for 72 hrs • Easy-to-use • High throughput • Reproducible for QC • Target cell specific • Run assay for 72 hrs • High throughput • Easy-to-use • Reproducible for QC Key Benefits
- 10. 1010 KILR™ Applications Antibody Dependent Cellular Cytotoxicity 1 0 -1 2 1 0 -1 1 1 0 -1 0 1 0 -9 1 0 -8 1 0 -7 1 0 -6 1 0 -5 0 1 0 2 0 3 0 4 0 E rb B 2 O v e r-e x p re s s in g C e ll P o o ls 4 :1 N K : targ et ce lls T ra s tu z u m a b [g /m L ]%lysis S /BHillSlope EC50 SKBR3 2.113 2.654e-009 MCF7 1.330 8.779e-009 NCI N87 0.7026 1.110e-008 1 2 .2 6 .1 1 0 .8 1 0 -12 1 0 -11 1 0 -10 1 0 -9 1 0 -8 1 0 -7 1 0 -6 1 0 -5 0 3 0 0 0 6 0 0 0 9 0 0 0 U 2 O S -E G F R e P L A n tib o d y [g /m L ] RLU S /B 8 .6 HillSlope EC50 Cetuximab 1.021 4.696e-009 Rituximab ~ 19.81 ~ 5.119e-011 -- ADCC Assay Verified with Marketed Drugs ADCC
- 11. 1111 Data provided courtesy of Amgen IC50 (ng/mL) Relative Potency R2 Ref. Std. 0.485 1 1 100% Ctrl. 0.519 0.961 0.999 60% 0.897 0.574 0.999 80% 0.581 0.83 0.996 130% 0.421 1.21 0.997 160% 0.324 1.64 0.998 KILR™ Applications ADCC for QC Lot Release Assays Key Benefits • True MOA of ADCC molecule • Accurate • Precise • Reproducible • Robust
- 12. 1212 KILR™ Applications Antibody Dependent Cellular Phagocytosis ADCP with Primary Macrophages Data provided courtesy of Janssen
- 13. 1313 KILR™ Applications Bispecific Antibody for T-cell Redirection Bi-specific Ab Plate Target Cells in 384-well plate Add non-activated primary T cells Add antibody Add KILR Detection reagent Read signal 41 hours 1hour Data provided courtesy of Janssen
- 14. 1414 Summary & Available KILR™ Assays • Highly-specific measure of target cell death in co-culture • Easy to use with simple add and read protocol • Highly sensitive detection of as few as 75 dead cells in a well • Broad applications for immuno-oncology drug development Product Part Number KILR ARH-77 Cell Line 97-1001C017 KILR H322 Cell Pool 97-1003P020 KILR NCI-N87 Cell Pool 97-1004P021 KILR Raji Cell Pool 97-1012P026 KILR Ramos Cell Pool 97-1006P022 KILR SKBR3 Cell Pool 97-1002P018 KILR A549 Cell Pool 97-1005P015 KILR SKOV3 Cell Pool 97-1007P023 KILR NCI-H292 Cell Pool 97-1008P024 KILR Daudi Cell Pool 97-1009P025 Product Part Number KILR Retroparticles for Suspension Cells (Neomycin) 97-0002 KILR Retroparticles for Adherent Cells (Neomycin) 97-0003 KILR Retroparticles for Adherent & Suspension Cells (Neomycin) 97-0004 Cell Lines Retrovirus To Make Your Own Cell Lines