Recent Advances in Immune Monitoring Presentation Slides
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The human immune system plays a pivotal role in conferring protective immunity to pathogenic microorganisms and cancer while its hyper-activation can result in serious inflammatory conditions such as autoimmunity. Immune monitoring is critical to determine the efficacy of therapeutic approaches such as the administration of a vaccine or to ascertain the safety of an administered agent such as a biologic that targets the immune system. Immune monitoring can involve the measurement of soluble immune mediators including cytokines and chemokines, as well as the analysis of phenotypic and functional status of immune cells. There is a plethora of platforms that offer highly sensitive and specific assays to monitor the immune system. These include simple immunoassays to complex multiplex assays that measure soluble biomarkers in various biological fluids and cell-based assays using a variety of platforms such as flow cytometry and enzyme-linked immunospot (ELISPOT) assay. In order to generate reliable immune monitoring data that will help us determine the safety and efficacy of the therapeutic intervention approach from a global study, highly standardized methods to collect, process and prepare patient samples need to be implemented. These slides focus on the recent advances in immune monitoring.
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Recent Advances in Immune Monitoring Presentation Slides
- 1. Public Recent Advances in Immune Monitoring | November 5, 20151Copyright © 2015 Covance. All Rights Reserved WEBINAR: RECENT ADVANCES IN IMMUNE MONITORING Sinnathamby Gomathinayagam, PhD
- 2. Public Recent Advances in Immune Monitoring | November 5, 20152 Why is Immune Monitoring Necessary? We monitor the immune system through a variety of means to: • Determine the efficacy of therapeutic approaches, such as the administration of a vaccine, or to • Ascertain the safety of an administered agent such as a biologic that targets the immune system.
- 3. Public Recent Advances in Immune Monitoring | November 5, 20153 ELISA SEREX ELISPOT Western Blot IHC Cytokine Release Assay Flow Cytometry PCR MHC Tetramers TCR Usage Proliferation Gene Array CTL Assay 2D Gel Electorphoresis Limiting Dilution Analysis SELDI DTH Mass Spectroscopy Commonly Used Techniques in Immune Profiling Whelan M et al., Personalized Medicine 2006 3(1), 79-88 SAFETY & EFFICACY ASSESSMENT
- 4. Public Recent Advances in Immune Monitoring | November 5, 20154 Biologics Targeting The Immune System Brennan et al., mAbs, 2010:2(3):233-255 Removab® Orencia® Stelara® Tysabri® Actemra® Ilaris® Humira® Remicade® Prolia® Soliris® OKT3® Enbrel® Simponi® Cimzia® Rituxan® Mabthera® Zevalin® Bexxar® Simulect® Amevive®
- 5. Public Recent Advances in Immune Monitoring | November 5, 20155 Immune Monitoring: “Cytokine Storm” Cytokine Storm in a Phase 1 Trial of the Anti-CD28 Monoclonal Antibody TGN1412 Ganesh Suntharalingam, F.R.C.A., Meghan R. Perry, M.R.C.P., Stephen Ward, F.R.C.A., Stephen J. Brett, M.D., Andrew Castell-Cortes, F.R.C.A., Michael D. Brunner, F.R.C.A., and Nicki Panoskaltsis, M.D., Ph.D. N Engl J Med 2006; 355:1018-28 The NEW ENGLAND JOURNAL of MEDICINE
- 6. Public Recent Advances in Immune Monitoring | November 5, 20156 Immune Monitoring: “Cytokine Storm”
- 7. Public Recent Advances in Immune Monitoring | November 5, 20157 Chemokine Panel Proinflammatory Panel Cytokine Panel IL-8 IFNy GM-CSF IP-10 IL-1B IL-12/IL-23 MCP-1 IL-2 IL-5 MCP-4 IL-6 IL-7 MDC IL-8 IL-15 MIP-1α IL-10 IL-16 MIP-1β IL-17A TNFβ VEGF C h e m o k in e P a n e l 0 .0 1 1 1 0 0 1 0 0 0 0 1 0 0 0 0 0 0 1 0 2 1 0 3 1 0 4 1 0 5 1 0 6 1 0 7 IL -8 (H A ) E o ta xin -3 IP -1 0 M C P -1 M C P -4 M D C M IP -1 M IP -1 T AR C C o n c [p g /m L ] ECLSignal 24-plex assay Immune Monitoring: “Cytokine Storm” MULTIPLEX CYTOKINE ASSAYS © Meso Scale Diagnostics , all rights reserved
- 8. Public Recent Advances in Immune Monitoring | November 5, 20158 • APRIL / TNFSF13 • BAFF / TNFSF13B • sCD30 / TNFRSF8 • sCD163 • Chitinase-3-like 1 • gp130 / sIL-6Rβ • IFN-α2 • IFN-β • IFN-γ • IL-2 • sIL-6Rα • IL-8 • IL-10 • IL-11 • IL-12 (p40) • IL-12 (p70) • IL-19 • IL-20 • IL-22 • IL-26 • IL-27 (p28) • IL-28A / IFN-λ2 • IL-29/IFN-λ1 • IL-32 • IL-34 • IL-35 • LIGHT / TNFSF14 • MMP-1 • MMP-2 • MMP-3 • Osteocalcin • Osteopontin • Pentraxin-3 • sTNF-R1 • sTNF-R2 • TSLP • TWEAK / TNFSF12 Bio-Rad Bio-Plex® Pro Human Inflammation Panel 1, 37-Plex Immune Monitoring: “Cytokine Storm” MULTIPLEX CYTOKINE ASSAYS Source: http://www.bio-rad.com/en-is/sku/171al001m-bio-plex-pro-human-inflammation-panel-1-37-plex
- 9. Public Recent Advances in Immune Monitoring | November 5, 20159 MULTI-ARRAY ® Meso Scale Diagnostics, LLC (Electro- chemiluminescence) Bio-Plex Bio-Rad (Luminex) A2® Beckman Coulter (Bead based chemiluminescence) FAST Quant Whatman Scleicher & Schuell BioScience ( Slide array system) IL-12p70 Manufacturer’s calibrator interval, ng/Lb 2500–0.6 4411–0.3 4800–6.5 50 000–12.2 Quantifiable interval, ng/Lc 2500–0.6 4411–0.27 577–7.1 625–2.4 Signal intervald ≅2 700 000–200 ≅23 000–30 ≅1000–25 ≅16 000–100 Mean CV within quantifiable intervale 9.60% 2.80% 8.70% 3.40% IL-2 Manufacturer’s calibrator interval, ng/L 2500–0.6 2161–0.1 6630–9.1 10 000–2.4 Quantifiable interval, ng/L 2500–2.4 540–2.1 245–9 10 000–2.4 Signal interval ≅600 000–200 ≅18 000–100 ≅4000–100 ≅2000–30 Mean CV within quantifiable interval 5.00% 5.90% 10.00% 3.20% IL-6 Manufacturer’s calibrator interval, ng/L 2500–0.6 2215–0.1 5200–7.1 10 000–2.4 Quantifiable interval, ng/L 2500–0.6 138–2.1 577–7.1 625–2.4 Signal interval ≅1 300 000–400 ≅6000–80 ≅1000–25 ≅16 000–100 Mean CV within quantifiable interval 4.70% 2.80% 8.70% 3.40% IL-10 Manufacturer’s calibrator interval, ng/L 2500–0.6 4311–0.3 4750–6.5 50 000–12.2 Quantifiable interval, ng/L 2500–0.6 269–1.05 175–6.5 50 000–195 Signal interval ≅500 000–300 ≅70 000–20 ≅3000–70 ≅1500–300 Mean CV within quantifiable interval 5% 8% 6% 4% IL-1β Manufacturer’s calibrator interval, ng/L 2500–0.6 3794–0.2 4840–6.6 10 000–2.4 Quantifiable interval, ng/L 2500–0.6 59–0.2 538–6.6 625–2.4 Signal interval ≅1 300 000–500 ≅9000–50 ≅3000–50 ≅20 000–400 Mean CV within quantifiable interval 6.60% 5.30% 8.40% 5.00% Calibrators Manufacturer’s recommended standard dilution factorf 1/4 Dilution 1/4 Dilution 1/3 Dilution 1/4 Dilution Calibration curve 7 Concentrations 8 Concentrations 7 Concentrations 7 Concentrations Comparison of the performance of 4 multiplex immunoassay platforms The MULTI-ARRAY and Bio-Plex assays had the best performance with the lowest limits of detection, and the MULTI-ARRAY system had the most linear signal output over the widest concentration range (105 to 106). Immune Monitoring: “Cytokine Storm” MULTIPLEX CYTOKINE ASSAYS Clin Chem. Author manuscript; available in PMC 2010 July 19.
- 10. Public Recent Advances in Immune Monitoring | November 5, 201510 Flow Cytometry in Immune Monitoring CHALLENGES FACED Limited amount of blood and other tissue material available from patients Need for standardized procedures across clinical trial sites PBMC isolation and cryopreservation Viability of samples upon thawing Experience of staff Whole blood vs. PBMC Phenotyping vs. Functional
- 11. Public Recent Advances in Immune Monitoring | November 5, 201511 Antigen-specific stimulation of T cells results in cytokine expression. Intracellular staining with IFNγ FITC/CD69 PE/CD8 PerCPCy5.5/CD3 APC by Ficoll or CPT-processed PBMC from one representative HIV+ donor. Flow Cytometry in Immune Monitoring Functional Assays Intracellular Cytokine Analysis Ruitenberg et al. BMC Immunology 2006 7:11 doi:10.1186/1471-2172-7-11
- 12. Public Recent Advances in Immune Monitoring | November 5, 201512 Intracellular Cytokine Analysis BioMed Research International Volume 2013, Article ID 726239, 11 pages http://dx.doi.org/10.1155/2013/726239
- 13. Public Recent Advances in Immune Monitoring | November 5, 201513 Hickling et al., 1998 Tetramer Analysis
- 14. Public Recent Advances in Immune Monitoring | November 5, 201514 Tetramer Analysis Immunology, 146, 11-22
- 15. Public Recent Advances in Immune Monitoring | November 5, 201515 Discovery of novel epitopes and validation Immune Monitoring 01 0.1 1 10 100 PHA g/ml 0.01 0.1 1 10 100 -200 0 200 400 600 PHA g/ml Numberofspots 0.01 0.1 1 10 100 -200 0 200 400 600 PHA g/ml Numberofspots 01 0.1 1 10 100 PHA g/ml 0.01 0.1 1 10 100 -200 0 200 400 600 PHA g/ml Numberofspots 0.01 0.1 1 10 100 -200 0 200 400 600 PHA g/ml Numberofspots Analysis of T cell activation ELISPOT analysis Source: Covance Translation and Biomarker Solutions
- 16. Public Recent Advances in Immune Monitoring | November 5, 201516 Cells 2015, 4, 84-95; doi:10.3390/cells4010084 Immune Monitoring: “Cytokine Storm” COMPARATIVE MULTI-DONOR STUDY OF IFNγ SECRETION AND EXPRESSION BY HUMAN PBMCS USING ELISPOT SIDE-BY-SIDE WITH ELISA AND FLOW CYTOMETRY ASSAYS MULTIPLEX CYTOKINE ASSAYS
- 17. Public Recent Advances in Immune Monitoring | November 5, 201517 Cells 2015, 4, 84-95; doi:10.3390/cells4010084 Immune Monitoring: “Cytokine Storm” COMPARATIVE MULTI-DONOR STUDY OF IFNγ SECRETION AND EXPRESSION BY HUMAN PBMCS USING ELISPOT SIDE-BY-SIDE WITH ELISA AND FLOW CYTOMETRY ASSAYS MULTIPLEX CYTOKINE ASSAYS
- 18. Public Recent Advances in Immune Monitoring | November 5, 201518 Bendall, S.C.et al., 2012; Trends in Immunology CyTOF® is a registered trademark of Fluidigm Canada Inc Time of Flight Antibodies labeled with elemental isotopes Nebulizer Analysis Cell 1 Cell 2 Cell 3 .FCS Quadrupole Integrate per cell Heavy (>100Da) Reporter atomic ions Light (<100Da) Overly abundant ions Flow Cytometry in Immune Monitoring MASS CYTOMETRY (CyTOF®)
- 19. Public Recent Advances in Immune Monitoring | November 5, 201519 Weighing the Pros and Cons of CyTOF® Pros 1. Traditional labelling techniques can be used 2. No autofluorescence; No spectral overlaps 3. Minimal to no color compensation 4. Designing panels is easier Cons 1. Slow acquisition of samples (>1000 events per second) 2. Much cleaner sample needed 3. Limited set of commercial antibodies 4. Complex data analysis 5. Cells cannot be recovered CyTOF® is a registered trademark of Fluidigm Canada Inc
- 20. Public Recent Advances in Immune Monitoring | November 5, 201520 Maecker and Harari Journal for ImmunoTherapy of Cancer (2015) 3:44 DOI 10.1186/s40425-015-0085-x Flow Cytometry in Immune Monitoring
- 21. Public Recent Advances in Immune Monitoring | November 5, 201521 MHC II MHC I CD4+ T CD8+ T Endosomal Cytosolic Antigen Processing and Presentation © 2009 QIAGEN, all rights reserved
- 22. Public Recent Advances in Immune Monitoring | November 5, 201522 T-cell Mediated Anti-Tumor Immunity
- 23. Public Recent Advances in Immune Monitoring | November 5, 201523 Tumorigenesis Isolation and identification of tumor specific epitopes Potential use as peptide vaccine Epitope Identification
- 24. Public Recent Advances in Immune Monitoring | November 5, 201524 Tumor cells express HLA-class I/peptide complexes on their surface Immunoprecipitation of HLA-class I/peptide complexes Isolation of HLA-class I bound peptides Identification of tumor specific epitopes Generation and validation of epitope specific CTLs Approach to Identify Tumor Epitopes Presented by HLA Class I Mass-spectrometry
- 25. Public Recent Advances in Immune Monitoring | November 5, 201525 Isolation Of “buffy coat” Stimulate with peptide Three cycles of restimulation with peptide Test effector function of CTLs by ELISpot assay (IFNg or Granzyme B) Generation of Peptide Specific CTLs in vitro Culture “buffy coat” derived cells
- 26. Public Recent Advances in Immune Monitoring | November 5, 201526 18-33 0C 37 0C Can the Identified Peptides with a Putative HLA-A2 Binding Motif be Bound by HLA-A2?
- 27. Public Recent Advances in Immune Monitoring | November 5, 201527 T2 T2+p13 IgG FITC W6/32 BB7.2 T2+Flu T2+L1 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count M1 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count Plot2: Not Gated 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count M1 M2 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count Plot2: Gated by : Gate 1 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count M1 M2 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count Plot2: Gated by : Gate 1 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count M1 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count Plot2: Not Gated 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count M1 M2 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count Plot2: Gated by : Gate 1 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count M1 M2 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count Plot2: Gated by : Gate 1 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count M1 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count Plot2: Not Gated 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count M1 M2 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count Plot2: Gated by : Gate 1 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count M1 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count Plot2: Not Gated 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count M1 M2 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count Plot2: Gated by : Gate 1 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count M1 M2 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count Plot2: Gated by : Gate 1 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count M1 M2 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count Plot2: Gated by : Gate 1 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count M1 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count Plot2: Not Gated 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count M1 M2 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count Plot2: Gated by : Gate 1 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count M1 M2 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count Plot2: Gated by : Gate 1 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count M1 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count Plot2: Not Gated 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count M1 M2 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count Plot2: Gated by : Gate 1 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count M1 M2 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count Plot2: Gated by : Gate 1 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count M1 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count Plot2: Not Gated 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count M1 M2 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count Plot2: Gated by : Gate 1 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count M1 M2 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count Plot2: Gated by : Gate 1 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count M1 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count Plot2: Not Gated 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count M1 M2 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count Plot2: Gated by : Gate 1 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count M1 M2 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count Plot2: Gated by : Gate 1 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count M1 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count Plot2: Not Gated 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count M1 M2 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count Plot2: Gated by : Gate 1 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count M1 M2 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count Plot2: Gated by : Gate 1 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count M1 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count Plot2: Not Gated 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count M1 M2 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count Plot2: Gated by : Gate 1 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count M1 M2 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count Plot2: Gated by : Gate 1 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count M1 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count Plot2: Not Gated 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count M1 M2 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count Plot2: Gated by : Gate 1 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count M1 M2 10e0 10e1 10e2 10e3 10e4 Green Fluorescence (GRN-HLog) 1007550250 Count Plot2: Gated by : Gate 1 T2+L2 T2+L3 T2+L4 T2+L5 T2+L6 T2+L7 T2+L8 T 2T 2 + p 1 3T 2 + F luT 2 + L 1T 2 + L 2T 2 + L 3T 2 + L 4T 2 + L 5T 2 + L 6T 2 + L 7T 2 + L 8 0 25 50 75 100 T 2T 2 + p 1 3T 2 + F luT 2 + L 1T 2 + L 2T 2 + L 3T 2 + L 4T 2 + L 5T 2 + L 6T 2 + L 7T 2 + L 8 0 100 200 300 400 W6/32 BB7.2 Peptide pulsed T2 MFI Stabilization of Surface HLA Class I by Synthetic Peptides Shetty, V., Sinnathamby, G., Nickens, Z., Shah, P., Hafner et al., J of Proteomics 74(5):728-43
- 28. Public Recent Advances in Immune Monitoring | November 5, 201528 Michael A. Morse et al. Clin Cancer Res 2011;17:3408-3419 Immune Monitoring Using ELISPOT analysis CLINICAL TRIALS
- 29. Public Recent Advances in Immune Monitoring | November 5, 201529 0.1 1 10 100 1000 0 25 50 75 100 200% 100% 50% [TA] %SpecificLysis PBMC phenotypingADCC assay https://en.wikipedia.org/wiki/Antibody-dependent_cell- mediated_cytotoxicity#/media/File:Antibody-dependent_Cellular_Cytotoxicity.svg Gated on CD3-CD56+ lymphs Antibody Dependent Cellular Cytotoxicity (ADCC)
- 30. Public Recent Advances in Immune Monitoring | November 5, 201530 0.001 0.01 0.1 1 -50 -25 0 25 50 75 100 125 150 1. 50% 2. 50% 3. 50% 1. 100% 2. 100% 3. 100% 1. 200% 2. 200% 3. 200% [Ab] g/ml %SpecificLysis 0.001 0.01 0.1 1 -50 -25 0 25 50 75 100 125 150 Assay 1 Assay 2 Assay 3 [Ab] g/ml %SpecificLysis Precision Accuracy Complement Dependent Cytotoxicity (CDC)
- 31. Public Recent Advances in Immune Monitoring | November 5, 201531 The Essential Role of Immune Monitoring ► Immune monitoring is a crucial component of clinical trials ► A variety of techniques can be employed to assess soluble mediators as well as cell associated biomarkers ► Multiplex immunoassays to determine the levels of soluble mediators and flow cytometry-based assays to determine the levels of activation markers on the surface of immune cells have played a major role in immune monitoring ► Employing the right technique at the right time is critical to determine the safety and efficacy of a therapeutic intervention ASSESSING SAFETY AND EFFICACY
- 32. Public Recent Advances in Immune Monitoring | November 5, 201532 Sinnathamby Gomathinayagam, PhD Sinnathamby.Gomathinayagam@covance.com
- 33. Public Recent Advances in Immune Monitoring | November 5, 201533 Covance Inc., headquartered in Princeton, NJ, is the drug development business of Laboratory Corporation of America® Holdings (LabCorp®). Covance is the marketing name for Covance Inc. and its subsidiaries around the world.