Measure modulations of immune checkpoint markers
Thermo Fisher Scientific offers a wide selection of ELISA and multiplex immunoassay solutions for immuno-oncology research that enable the measurement of soluble immune checkpoint molecules and other immune-related targets in serum, plasma, and culture supernatant for human and mouse.
Overview of immuno-oncology and immune checkpoint markers
Immuno-oncology is a type of immunotherapy approach that uses the body’s immune system to help battle cancer—a different approach that goes beyond traditional methods such as surgery, chemotherapy, and radiation.
Many recent drug development strategies have been to target immune checkpoint markers to enhance anti-tumor immunity. Immune checkpoints are cell pathways important in maintaining a normal immune response and protecting tissues from damage when the immune system is activated (Figure 1). Cancer cells dysregulate immune checkpoints and use them as a mechanism of immune resistance. For example, PD-1 masks cancer cells from T-cell recognition, thereby preventing the attack by T cells.
Cell signaling is a complex communication process to coordinate cell activities. Cell surface receptors respond to the microenvironment and trigger signal transduction cascades within the cell. The targets of such signaling pathways commonly include transcription factors that function to regulate gene expression. Errors in signaling interactions and cellular information processes are responsible for many diseases such as cancer and autoimmunity.
Cytokines are immune cell messengers that stimulate immune effector cells and stromal cells in tumors and also increase the recognition of tumor cells by cytotoxic effector cells. Investigations using animal models have revealed that cytokines display anti-tumor activity, resulting in an increased interest in the use of cytokines as a basis for cancer therapy. Many cytokines, such as GM-CSF, IL-7, IL-12, IL-15, IL-18, and IL-21, are in clinical trials for patients with advanced cancers. Pre-clinical research has focused on being able to neutralize cytokines that act to suppress the immune response, for example IL-10 and TGF-β. Adoptive cell therapy is an approach that makes modification to T cells to focus them on killing tumor cells, while controlling the cytokine response for optimal effect.
Figure 1. The cancer immunity cycle. Immune checkpoint inhibitors are targets for biotherapeutic antibodies.
Immuno-oncology ELISA kits
Detecting and measuring immune checkpoint markers could be a promising pathway to better develop treatment strategies. For example, CTLA-4 plays a role during the priming of a T cell by an antigen-presenting cell. Blocking of the CTLA-4 receptor by an antibody allows T cell activation, resulting in an anticancer immune response. Correlation has been observed with positive health benefits and elevated levels of quantified soluble CTLA-4 (sCTLA4) concentrations by ELISA in antibody-therapy treated subjects (Figure 2).
Whether you are studying CTLA-4 or other immune checkpoint markers, our ELISA kits provide a specific and sensitive method to measure and monitor these biomarkers in serum, plasma, or cell culture supernatants. Invitrogen ELISA kits for popular targets such as PD-1, CD80, TIM3 etc. are listed in Table 1.
Search immuno-oncology ELISA kits
Learn more about ELISA kits and components
Popular immuno-oncology protein targets and ELISA performance data
Table 1. View our ELISA kits for the following popular targets:
Figure 2. Serum CTLA-4 levels in melanoma patient samples. Statistical analysis following ipilimumab (CTLA-4 antibody drug) therapy shows correlation between responders and higher concentration levels of CTLA-4 in serum samples of melanoma patients using CTLA-4 (Soluble) Human ELISA Kit.
Reprinted from: Leung A.M., et al. Clinical benefit from ipilimumab therapy in melanoma patients may be associated with serum CTLA4 levels. Front Oncol, 2014. 4: 110.
Immuno-oncology ProQuantum high sensitivity immunoassays
ProQuantum high-sensitivity immunoassays are designed for easy-to-run, high-performance, protein detection with no proprietary instrument to purchase. Utilizing proximity-based amplification technology, ProQuantum assays offer the analyte specificity of antibody-antigen binding with the signal detection and amplification capabilities of qPCR. ProQuantum assays can also typically detect lower levels of protein with lower sample consumption than traditional methods. This can be particularly important when handling very limited volumes of precious samples from blood or tumor extracts. Limited sample volumes also present a challenge to detect very low levels of protein, but our ProQuantum technology solves that by amplifying the signal beyond the capability of other available methods.
See our selection of sensitive assays for cytokines, which play an important role in immuno-oncology and checkpoint modulation (Table 2). Click below to see our complete selection of assays.
Find cytokine-related ProQuantum assays
Learn more about how the ProQuantum immunoassays work
Read BioProbes Journal article: Introducing ProQuantum High-Sensitivity Immunoassays—The new generation of target-specific protein quantitation
Table 2. Examples of ProQuantum immunoassays for immuno-oncology research*.
| Cat. No. | Cytokine | Primary cell source | Primary target cell | Biological activity |
|---|---|---|---|---|
| A428915 | GM-CSF | T cells, macrophages, endothelial cells, fibroblasts, mast cells | Bone marrow, progenitor cells, DC, macrophages NKT cells | Promotes antigen presentation, T cell homeostasis, hematopoietic cell growth factor |
| A35574 | IL-1 | Monocytes, macrophages, fibroblasts, epithelial cells, endothelial cells, astrocytes | T cells, B cells, endothelial cells, hypothalamus, liver | Co-stimulation, cell activation, inflammation, fever, acute phase reactant |
| A35603 | IL-2 | T cells, NK cells | T cells, NK cells, B cells, monocytes | Cell growth/activation |
| IL-3 | T cells | Bone marrow progenitor cells | Cell growth and differentiation | |
| A35587 | IL-4 | T cells | T cells, B cells | Th2 differentiation, cell growth/activation, IgE isotype switching |
| A35588 | IL-5 | T cells | B cells, eosinophils | Cell growth/activation |
| A35573 | IL-6 | T cells, macrophages, fibroblasts | T cells, B cells, liver | Co-stimulation, cell growth/activation, acute phase reactant |
| A35589 | IL-7 | Fibroblasts, bone marrow stromal cells | Immature lymphoid, progenitors | T cell survival, proliferation, homeostasis, B cell development |
| A35575 | IL-8 | Macrophages, epithelial cells, platelets | Neutrophils | Activation, chemotaxis |
| A35590 | IL-10 | Th2 T cells | Macrophages, T cells | Inhibits antigen-presenting cells, inhibits cytokine production |
| A35577 | IL-12 | Macrophages, NK cells | T cells | Th1 differentiation |
| IL-15 | Monocytes | T cells, NK cells | Cell growth/activation, NK cell development, blocks apoptosis | |
| A35613 | IL-18 | Macrophages | T cells, NK cells, B cells | Cell growth/activation, inflammation |
| A35593 | IL-21 | CD4+ T cells, NKT cells | NK cells, T cells, B cells | Cell growth/activation, control of allergic responses and viral infections |
| IL-23 | Antigen-presenting cells | T cells, NK cells, DC | Chronic inflammation, promotes Th17 cells | |
| A42897 | IFN-α | Plasmacytoid DC, NK cells, T cells, B cells, macrophages, fibroblasts, endothelial cells, osteoblasts | Macrophages, NK cells | Anti-viral, enhances MHC expression |
| A35576 | IFN-γ | T cells, NK cells, NKT cells | Monocytes, macrophages, endothelial cells, tissue cells | Cell growth/activation, enhances MHC expression |
| A42898 | TGF-β | T cells, macrophages | T cells | Inhibits cell growth/activation |
| A35601 | TNF- α | Macrophages, T cells | T cells, B cells, endothelial cells, hypothalamus, liver | Co-stimulation, cell activation, inflammation, fever, acute phase reactant |
* S. Lee and K. Margolin. Cytokines in Cancer Immunotherapy. Cancers (Basel), 2011. 3(4): 3856-93.
Immuno-oncology ProcartaPlex multiplex immunoassays
Invitrogen ProcartaPlex Immuno-Oncology Checkpoint Panels allow for the simultaneous detection of up to 38 soluble immune checkpoint molecules and help provide a more comprehensive picture of cancer immunity.
Several checkpoint inhibitors were detected in tumor tissue extracts from mouse using the ProcartaPlex Mouse Immuno-Oncology Checkpoint Panel, 4plex and ProcartaPlex Mouse Immuno-Oncology Checkpoint Panel 2, 7plex (Figure 3). We have also measured checkpoint markers in melanoma patient samples using the ProcartaPlex Human Immuno-Oncology Checkpoint Panel 1, 14plex and ProcartaPlex Human Immuno-Oncology Checkpoint Panel 2, 14plex (Figure 4).
Our Immuno-Oncology Checkpoint panels have been specifically designed and optimized for cerebral spinal fluid, serum, and plasma samples (Table 3). Use preconfigured panels below or select from single targets to create your own custom panel to study either human or mouse immune checkpoint biomarkers.
ProcartaPlex Panel Configurator
Learn more about ProcartaPlex multiplex immunoassays
Watch these on-demand webinars:
- Jun 2021 Panel Presentation: xMAP® INTELLIFLEX and Immunoassay Innovations: Tools to advance immuno-oncology research
Presented by Wilco de Jager, PhD, Sr. Field Applications Scientist, Luminex and Sigrun Badrnya, Product Development Manager, Thermo Fisher Scientific - Sep 2018 Poster Presentation: Development of Multiplex xMAP® Technology-Based Assays for Simultaneous Detection of Soluble Checkpoint Molecules Involved in Anti-Cancer Immune Response
Presented by Susanne Oehler, PhD, Senior Manager, Research and Development, Antibodies and Immunoassays, Thermo Fisher Scientific
Preconfigured immuno-oncology multiplex immunoassay panels and performance data
Figure 3. Detection of multiple immune checkpoint inhibitors in mouse samples. The ProcartaPlex Mouse Immuno-Oncology Checkpoint Panel, 4plex (top) and ProcartaPlex Mouse Immuno-Oncology Checkpoint Panel 2, 7plex (bottom) were tested using homogenized MC38 or TC-1 derived tumor tissue extract from C57BI/6 mice. The tumor tissue was removed at different time points post-treatment. Tissue extracts were homogenized in RIPA Buffer using a proteinase inhibitor. Untreated mice served as control. Differences were observed between the treated vs. control groups, suggesting role of these proteins in tumor progression. Data was provided by our collaborator.
Figure 4. Serum levels of checkpoint markers in melanoma patient samples. The ProcartaPlex Human Immuno-Oncology Checkpoint Panel 1, 14plex and ProcartaPlex Human Immuno-Oncology Checkpoint Panel 2, 14plex were used to measure various checkpoint markers in melanoma patient samples. Results are shown as the mean of ungrouped human samples for all targets of both the panels. Used with permission from Exner R., Sachet M., Arnold T., et al. Prognostic value of HMGB1 in early breast cancer patients under neoadjuvant chemotherapy. Cancer Med, 2016. 5(9): 2350-8.
Table 3. Preconfigured ProcartaPlex multiplex immunoassay panels for immuno-oncology.
| Human multiplex immune checkpoint panels | ||
| Product name | Size | Cat. No. |
| ProcartaPlex Human Immuno-Oncology Checkpoint Panel 1, 14plex Target list [bead region]: Immune stimulatory: CD27 [27], CD28 [15], CD137 (4-1BB) [26], GITR [57], HVEM [36] Immune inhibitory: BTLA [52], CD80 [61], CD152 (CTLA4) [33], IDO [46], LAG-3 [47], PD-1 [65], PD-L1 [66], PD-L2 [67], TIM-3 [14] | 96 tests | |
| ProcartaPlex Human Immuno-Oncology Checkpoint Panel 2, 14plex Target list [bead region]: Activating: MICA [18], MICB [21], Perforin [53], ULBP-1 [73], ULBP-3 [77], ULBP-4 [78] Inhibitory: Arginase-1 [51], CD73 (NT5E) [30], CD96 (Tactile) [35], E-Cadherin [44], Nectin-2 [29], PVR [56], Siglec-7 [12], Siglec-9 [13] | 96 tests | EPX140-15815-901 |
| Mouse multiplex immune checkpoint panels | ||
| ProcartaPlex Mouse Immuno-Oncology Checkpoint Panel, 4plex Target list [bead region]: BTLA [61], CD27 [75], LAG-3 [44], TIM-3 [46] | 96 tests | EPX040-20830-901 |
| ProcartaPlex Mouse Immuno-Oncology Checkpoint Panel 2, 7plex Target list [bead region]: CD137L (4-1BBL) [74], CD152 (CTLA4) [77], CD276 (B7-H3) [67], CD80 [15], PD-1 [73], PD-L1 [29], PD-L2 [53] | 96 tests | EPX070-20835-901 |
Multiplex gene expression and protein assays—immuno-oncology research
QuantiGene RNA gene expression assays provide a fast and high-throughput solution for multiplexed gene expression quantitation, with simultaneous measurement of up to 80 genes of interest in a single well of a 96- or 384-well plate. The QuantiGene Plex assay is based on hybridization and incorporates branched DNA (bDNA) technology, which uses signal amplification rather than target amplification for direct measurement of RNA transcripts. The assay is run on the Luminex platform, has a simple workflow, and does not require RNA purification. These features allow the user to merge the QuantiGene workflow for gene expression profiling with the ProcartaPlex workflow for protein quantitation using the same sample (Figure 5).
Learn more about QuantiGene RNA Assays for Gene Expression Profiling
Read our publication: Multiplexing protein and gene level measurements on a single Luminex platform. Methods, 2019. 158: 27-32.
Figure 5. Combined workflow for QuantiGene gene expression and ProcartaPlex protein quantitation assays.
Researchers can now gain deeper insights through the preconfigured panels such as the QuantiGene Plex Human Immune Response Panel, 80-plex, which simultaneously analyzes 80 cytokine, chemokine, and growth factor targets. These targets are complementary and correlate to the ProcartaPlex Human Immune Response Panel, 80plex allowing maximum data per sample using the same instrument (Figure 6). Full data, along with the target list and correlation analysis can be found here.
Additional panels are also available for curated research into immune profiling and cancer biomarkers across human and mice samples (Table 4).
Figure 6. Correlation of gene (RNA) vs. protein expression in LPS-stimulated PBMCs. PBMC were stimulated with 10 µg/mL of Lipopolysaccharide (LPS). Correlation of RNA and protein expression of ENA (CXCL5), GRO-alpha (CXCL1), MCP-3 (CCL7) and BLC (CXCL13) were measured at 48h post LPS-stimulation. Raw MFI data from the QuantiGene Plex Human Immune Response Panel, 80-plex was normalized to the housekeeping control, PPIB. Protein data was acquired using the complementary ProcartaPlex Human Immune Response Panel, 80plex. Data is displayed as normalized gene expression (RNA) and total amounts of protein (pg/mL) for unstimulated and LPS-stimulated samples at the 48h timepoint. RNA expression is represented by lines and protein expression by bars.
Table 4. Immune Response and Cancer Profiling Gene Expression Panels.
| Product name | Size | Cat. No. |
QuantiGene Plex Human Immune Response Panel, 80-plex Targets: CCL1, CCL13, CCL17 ,CCL19, CCL2, CCL21, CCL22, CCL23, CCL24, CCL25, CCL26, CCL4, CCL7, CCL8, CD40LG, CSF1, CSF2, CSF3, CX3CL1,CXCL1, CXCL11, CXCL13, CXCL2, CXCL5, CXCL6, CXCL9, CXCR3, FGF2, GZMA, GZMB, HGF, IFNA1, IFNG, IL10, IL12A, IL12B, IL13, IL15, IL16, IL17A, IL18, IL1A, IL1B, IL2, IL20, IL21, IL22, IL23A, IL27, IL2RA, IL3, IL31, IL34, IL37, IL4, IL5, IL6, IL8, IL9, KITLG, LGALS3, LIF, LTA, MIF, NGF, PTX3, TNF, TNFRSF12A, TNFRSF1B, TNFRSF8, TNFSF10, TNFSF13, TNFSF13B, TREM1, TSLP, VEGFA, PPIB, HPRT1, GAPDH, GUSB | 1 plate | QGP-180-10080 |
3 plates | ||
| 10 plates | ||
QuantiGene Plex Mouse Immune Response Panel, 80-plex Targets: Il2ra, Il2, Il6, Ifng, Tnf, Il5, Il1a, Csf2, Il4, Il10, Il18, Vegfa, Il17a, Il1b, Il12a, Ccl2, Ccl7, Ccl11, Ccl5, Cxcl11, Il6ra, Ccl3, Ccl4, Il13, Il23a, Cxcr3, Il22, Il15, Il27, Il33, Ifna1, Il28a, Il31, Cxcl1, Cxcl2, Csf3, Il3, Lep, Tnfsf11, Csf1, Lif, Il9, Btc, Cxcl5, Il25, Il1rl1, Il19, Cd27, Kdr, Ccl19, Cxcl16, Il16, Ccl22, Ccl12, Gzmb, Ccl27a, Ccl24, Ccl17, Ccl25, Cxcl13, Il7r, Il7, Tslp, Tnfsf13b, Il12b, Il21, Gzma, Cd274, Ctla4, Cxcl9, Havcr2, Lag3, Ifna2, Ifnb1, Hgf, Tnfrsf12a, Ppib, Hprt, Gapdh, Gusb | 1 plate | |
3 plates | ||
10 plates | ||
QuantiGene Plex Human PanCancer Panel, 80-plex Targets: AGER, ARG1, AXL, BDNF, BTLA, CALR, CD27, CD274, CD276, CD28, CD36, CD47, CD48, CD80, CD96, CDH1, CSF3, CTLA4, CXCL8, DKK1, EGF, EPCAM, FGF2, GAPDH, GAS6, GPC1, GUSB, HAVCR2, HGF, HMGB1, HPRT1, HSP90AA1, HSPA4, HSPB2, HSPD1, ICOSLG, IDO1, IGF2, KITLG, LAG3, LGALS9, LIF, MBL2, MDK, MERTK, MICA, MICB, NCR3LG1, NECTIN2, NGF, NT5E, NTRK2, OLR1, PDCD1LG2, PECAM1, PGF, PLAUR, PPIB, PRF1, PVR, RAET1E, S100A8, S100A9, SERPINE1, SIGLEC7, SIGLEC9, SPARC, SPATA2, TIMD4, TNFRSF14, TNFRSF18, TNFRSF4, TNFRSF9, TRIM8, TYRO3, ULBP1, ULBP3, VEGFA, VEGFD, VSIR | 1 plate | |
3 plates | ||
10 plates | ||
QuantiGene Plex Mouse PanCancer Panel, 80-plex Targets: Ager, Arg1, Axl, Bdnf, Btla, Calr, Cd27, Cd274, Cd276, Cd28, Cd36, Cd47, Cd48, Cd80, Cd96, Cdh1, Csf3, Ctla4, Cxcl1, Dkk1, Egf, Epcam, Fgf2, Gapdh, Gas6, Gpc1, Gusb, Havcr2, Hgf, Hmgb1, Hprt, Hsp90Aa1, Hspa4, Hspb2, Hspd1, Icosl, Ido1, Igf2, Inhca , Kitl, Lag3, Lgals9, Lif, Mbl2, Mdk, Mertk, Mill2, Ncr1, Nectin2, Ngf, Nt5E, Ntrk2, Olr1, Pdcd1Lg2, Pecam1, Pgf, Plaur, Ppib, Prf1, Pvr, Raet1E, S100A8, S100A9, Serpine1, Siglece, Siglech, Sparc, Spata2, Timd4, Tnfrsf14 , Tnfrsf18, Tnfrsf4, Tnfrsf9, Trim8, Tyro3, Ulbp1, Ulbp3, Vegfa, Vegfd, Vsir | 1 plate | |
3 plates | ||
10 plates |
Overview of immuno-oncology and immune checkpoint markers
Immuno-oncology is a type of immunotherapy approach that uses the body’s immune system to help battle cancer—a different approach that goes beyond traditional methods such as surgery, chemotherapy, and radiation.
Many recent drug development strategies have been to target immune checkpoint markers to enhance anti-tumor immunity. Immune checkpoints are cell pathways important in maintaining a normal immune response and protecting tissues from damage when the immune system is activated (Figure 1). Cancer cells dysregulate immune checkpoints and use them as a mechanism of immune resistance. For example, PD-1 masks cancer cells from T-cell recognition, thereby preventing the attack by T cells.
Cell signaling is a complex communication process to coordinate cell activities. Cell surface receptors respond to the microenvironment and trigger signal transduction cascades within the cell. The targets of such signaling pathways commonly include transcription factors that function to regulate gene expression. Errors in signaling interactions and cellular information processes are responsible for many diseases such as cancer and autoimmunity.
Cytokines are immune cell messengers that stimulate immune effector cells and stromal cells in tumors and also increase the recognition of tumor cells by cytotoxic effector cells. Investigations using animal models have revealed that cytokines display anti-tumor activity, resulting in an increased interest in the use of cytokines as a basis for cancer therapy. Many cytokines, such as GM-CSF, IL-7, IL-12, IL-15, IL-18, and IL-21, are in clinical trials for patients with advanced cancers. Pre-clinical research has focused on being able to neutralize cytokines that act to suppress the immune response, for example IL-10 and TGF-β. Adoptive cell therapy is an approach that makes modification to T cells to focus them on killing tumor cells, while controlling the cytokine response for optimal effect.
Figure 1. The cancer immunity cycle. Immune checkpoint inhibitors are targets for biotherapeutic antibodies.
Immuno-oncology ELISA kits
Detecting and measuring immune checkpoint markers could be a promising pathway to better develop treatment strategies. For example, CTLA-4 plays a role during the priming of a T cell by an antigen-presenting cell. Blocking of the CTLA-4 receptor by an antibody allows T cell activation, resulting in an anticancer immune response. Correlation has been observed with positive health benefits and elevated levels of quantified soluble CTLA-4 (sCTLA4) concentrations by ELISA in antibody-therapy treated subjects (Figure 2).
Whether you are studying CTLA-4 or other immune checkpoint markers, our ELISA kits provide a specific and sensitive method to measure and monitor these biomarkers in serum, plasma, or cell culture supernatants. Invitrogen ELISA kits for popular targets such as PD-1, CD80, TIM3 etc. are listed in Table 1.
Search immuno-oncology ELISA kits
Learn more about ELISA kits and components
Popular immuno-oncology protein targets and ELISA performance data
Table 1. View our ELISA kits for the following popular targets:
Figure 2. Serum CTLA-4 levels in melanoma patient samples. Statistical analysis following ipilimumab (CTLA-4 antibody drug) therapy shows correlation between responders and higher concentration levels of CTLA-4 in serum samples of melanoma patients using CTLA-4 (Soluble) Human ELISA Kit.
Reprinted from: Leung A.M., et al. Clinical benefit from ipilimumab therapy in melanoma patients may be associated with serum CTLA4 levels. Front Oncol, 2014. 4: 110.
Immuno-oncology ProQuantum high sensitivity immunoassays
ProQuantum high-sensitivity immunoassays are designed for easy-to-run, high-performance, protein detection with no proprietary instrument to purchase. Utilizing proximity-based amplification technology, ProQuantum assays offer the analyte specificity of antibody-antigen binding with the signal detection and amplification capabilities of qPCR. ProQuantum assays can also typically detect lower levels of protein with lower sample consumption than traditional methods. This can be particularly important when handling very limited volumes of precious samples from blood or tumor extracts. Limited sample volumes also present a challenge to detect very low levels of protein, but our ProQuantum technology solves that by amplifying the signal beyond the capability of other available methods.
See our selection of sensitive assays for cytokines, which play an important role in immuno-oncology and checkpoint modulation (Table 2). Click below to see our complete selection of assays.
Find cytokine-related ProQuantum assays
Learn more about how the ProQuantum immunoassays work
Read BioProbes Journal article: Introducing ProQuantum High-Sensitivity Immunoassays—The new generation of target-specific protein quantitation
Table 2. Examples of ProQuantum immunoassays for immuno-oncology research*.
| Cat. No. | Cytokine | Primary cell source | Primary target cell | Biological activity |
|---|---|---|---|---|
| A428915 | GM-CSF | T cells, macrophages, endothelial cells, fibroblasts, mast cells | Bone marrow, progenitor cells, DC, macrophages NKT cells | Promotes antigen presentation, T cell homeostasis, hematopoietic cell growth factor |
| A35574 | IL-1 | Monocytes, macrophages, fibroblasts, epithelial cells, endothelial cells, astrocytes | T cells, B cells, endothelial cells, hypothalamus, liver | Co-stimulation, cell activation, inflammation, fever, acute phase reactant |
| A35603 | IL-2 | T cells, NK cells | T cells, NK cells, B cells, monocytes | Cell growth/activation |
| IL-3 | T cells | Bone marrow progenitor cells | Cell growth and differentiation | |
| A35587 | IL-4 | T cells | T cells, B cells | Th2 differentiation, cell growth/activation, IgE isotype switching |
| A35588 | IL-5 | T cells | B cells, eosinophils | Cell growth/activation |
| A35573 | IL-6 | T cells, macrophages, fibroblasts | T cells, B cells, liver | Co-stimulation, cell growth/activation, acute phase reactant |
| A35589 | IL-7 | Fibroblasts, bone marrow stromal cells | Immature lymphoid, progenitors | T cell survival, proliferation, homeostasis, B cell development |
| A35575 | IL-8 | Macrophages, epithelial cells, platelets | Neutrophils | Activation, chemotaxis |
| A35590 | IL-10 | Th2 T cells | Macrophages, T cells | Inhibits antigen-presenting cells, inhibits cytokine production |
| A35577 | IL-12 | Macrophages, NK cells | T cells | Th1 differentiation |
| IL-15 | Monocytes | T cells, NK cells | Cell growth/activation, NK cell development, blocks apoptosis | |
| A35613 | IL-18 | Macrophages | T cells, NK cells, B cells | Cell growth/activation, inflammation |
| A35593 | IL-21 | CD4+ T cells, NKT cells | NK cells, T cells, B cells | Cell growth/activation, control of allergic responses and viral infections |
| IL-23 | Antigen-presenting cells | T cells, NK cells, DC | Chronic inflammation, promotes Th17 cells | |
| A42897 | IFN-α | Plasmacytoid DC, NK cells, T cells, B cells, macrophages, fibroblasts, endothelial cells, osteoblasts | Macrophages, NK cells | Anti-viral, enhances MHC expression |
| A35576 | IFN-γ | T cells, NK cells, NKT cells | Monocytes, macrophages, endothelial cells, tissue cells | Cell growth/activation, enhances MHC expression |
| A42898 | TGF-β | T cells, macrophages | T cells | Inhibits cell growth/activation |
| A35601 | TNF- α | Macrophages, T cells | T cells, B cells, endothelial cells, hypothalamus, liver | Co-stimulation, cell activation, inflammation, fever, acute phase reactant |
* S. Lee and K. Margolin. Cytokines in Cancer Immunotherapy. Cancers (Basel), 2011. 3(4): 3856-93.
Immuno-oncology ProcartaPlex multiplex immunoassays
Invitrogen ProcartaPlex Immuno-Oncology Checkpoint Panels allow for the simultaneous detection of up to 38 soluble immune checkpoint molecules and help provide a more comprehensive picture of cancer immunity.
Several checkpoint inhibitors were detected in tumor tissue extracts from mouse using the ProcartaPlex Mouse Immuno-Oncology Checkpoint Panel, 4plex and ProcartaPlex Mouse Immuno-Oncology Checkpoint Panel 2, 7plex (Figure 3). We have also measured checkpoint markers in melanoma patient samples using the ProcartaPlex Human Immuno-Oncology Checkpoint Panel 1, 14plex and ProcartaPlex Human Immuno-Oncology Checkpoint Panel 2, 14plex (Figure 4).
Our Immuno-Oncology Checkpoint panels have been specifically designed and optimized for cerebral spinal fluid, serum, and plasma samples (Table 3). Use preconfigured panels below or select from single targets to create your own custom panel to study either human or mouse immune checkpoint biomarkers.
ProcartaPlex Panel Configurator
Learn more about ProcartaPlex multiplex immunoassays
Watch these on-demand webinars:
- Jun 2021 Panel Presentation: xMAP® INTELLIFLEX and Immunoassay Innovations: Tools to advance immuno-oncology research
Presented by Wilco de Jager, PhD, Sr. Field Applications Scientist, Luminex and Sigrun Badrnya, Product Development Manager, Thermo Fisher Scientific - Sep 2018 Poster Presentation: Development of Multiplex xMAP® Technology-Based Assays for Simultaneous Detection of Soluble Checkpoint Molecules Involved in Anti-Cancer Immune Response
Presented by Susanne Oehler, PhD, Senior Manager, Research and Development, Antibodies and Immunoassays, Thermo Fisher Scientific
Preconfigured immuno-oncology multiplex immunoassay panels and performance data
Figure 3. Detection of multiple immune checkpoint inhibitors in mouse samples. The ProcartaPlex Mouse Immuno-Oncology Checkpoint Panel, 4plex (top) and ProcartaPlex Mouse Immuno-Oncology Checkpoint Panel 2, 7plex (bottom) were tested using homogenized MC38 or TC-1 derived tumor tissue extract from C57BI/6 mice. The tumor tissue was removed at different time points post-treatment. Tissue extracts were homogenized in RIPA Buffer using a proteinase inhibitor. Untreated mice served as control. Differences were observed between the treated vs. control groups, suggesting role of these proteins in tumor progression. Data was provided by our collaborator.
Figure 4. Serum levels of checkpoint markers in melanoma patient samples. The ProcartaPlex Human Immuno-Oncology Checkpoint Panel 1, 14plex and ProcartaPlex Human Immuno-Oncology Checkpoint Panel 2, 14plex were used to measure various checkpoint markers in melanoma patient samples. Results are shown as the mean of ungrouped human samples for all targets of both the panels. Used with permission from Exner R., Sachet M., Arnold T., et al. Prognostic value of HMGB1 in early breast cancer patients under neoadjuvant chemotherapy. Cancer Med, 2016. 5(9): 2350-8.
Table 3. Preconfigured ProcartaPlex multiplex immunoassay panels for immuno-oncology.
| Human multiplex immune checkpoint panels | ||
| Product name | Size | Cat. No. |
| ProcartaPlex Human Immuno-Oncology Checkpoint Panel 1, 14plex Target list [bead region]: Immune stimulatory: CD27 [27], CD28 [15], CD137 (4-1BB) [26], GITR [57], HVEM [36] Immune inhibitory: BTLA [52], CD80 [61], CD152 (CTLA4) [33], IDO [46], LAG-3 [47], PD-1 [65], PD-L1 [66], PD-L2 [67], TIM-3 [14] | 96 tests | |
| ProcartaPlex Human Immuno-Oncology Checkpoint Panel 2, 14plex Target list [bead region]: Activating: MICA [18], MICB [21], Perforin [53], ULBP-1 [73], ULBP-3 [77], ULBP-4 [78] Inhibitory: Arginase-1 [51], CD73 (NT5E) [30], CD96 (Tactile) [35], E-Cadherin [44], Nectin-2 [29], PVR [56], Siglec-7 [12], Siglec-9 [13] | 96 tests | EPX140-15815-901 |
| Mouse multiplex immune checkpoint panels | ||
| ProcartaPlex Mouse Immuno-Oncology Checkpoint Panel, 4plex Target list [bead region]: BTLA [61], CD27 [75], LAG-3 [44], TIM-3 [46] | 96 tests | EPX040-20830-901 |
| ProcartaPlex Mouse Immuno-Oncology Checkpoint Panel 2, 7plex Target list [bead region]: CD137L (4-1BBL) [74], CD152 (CTLA4) [77], CD276 (B7-H3) [67], CD80 [15], PD-1 [73], PD-L1 [29], PD-L2 [53] | 96 tests | EPX070-20835-901 |
Multiplex gene expression and protein assays—immuno-oncology research
QuantiGene RNA gene expression assays provide a fast and high-throughput solution for multiplexed gene expression quantitation, with simultaneous measurement of up to 80 genes of interest in a single well of a 96- or 384-well plate. The QuantiGene Plex assay is based on hybridization and incorporates branched DNA (bDNA) technology, which uses signal amplification rather than target amplification for direct measurement of RNA transcripts. The assay is run on the Luminex platform, has a simple workflow, and does not require RNA purification. These features allow the user to merge the QuantiGene workflow for gene expression profiling with the ProcartaPlex workflow for protein quantitation using the same sample (Figure 5).
Learn more about QuantiGene RNA Assays for Gene Expression Profiling
Read our publication: Multiplexing protein and gene level measurements on a single Luminex platform. Methods, 2019. 158: 27-32.
Figure 5. Combined workflow for QuantiGene gene expression and ProcartaPlex protein quantitation assays.
Researchers can now gain deeper insights through the preconfigured panels such as the QuantiGene Plex Human Immune Response Panel, 80-plex, which simultaneously analyzes 80 cytokine, chemokine, and growth factor targets. These targets are complementary and correlate to the ProcartaPlex Human Immune Response Panel, 80plex allowing maximum data per sample using the same instrument (Figure 6). Full data, along with the target list and correlation analysis can be found here.
Additional panels are also available for curated research into immune profiling and cancer biomarkers across human and mice samples (Table 4).
Figure 6. Correlation of gene (RNA) vs. protein expression in LPS-stimulated PBMCs. PBMC were stimulated with 10 µg/mL of Lipopolysaccharide (LPS). Correlation of RNA and protein expression of ENA (CXCL5), GRO-alpha (CXCL1), MCP-3 (CCL7) and BLC (CXCL13) were measured at 48h post LPS-stimulation. Raw MFI data from the QuantiGene Plex Human Immune Response Panel, 80-plex was normalized to the housekeeping control, PPIB. Protein data was acquired using the complementary ProcartaPlex Human Immune Response Panel, 80plex. Data is displayed as normalized gene expression (RNA) and total amounts of protein (pg/mL) for unstimulated and LPS-stimulated samples at the 48h timepoint. RNA expression is represented by lines and protein expression by bars.
Table 4. Immune Response and Cancer Profiling Gene Expression Panels.
| Product name | Size | Cat. No. |
QuantiGene Plex Human Immune Response Panel, 80-plex Targets: CCL1, CCL13, CCL17 ,CCL19, CCL2, CCL21, CCL22, CCL23, CCL24, CCL25, CCL26, CCL4, CCL7, CCL8, CD40LG, CSF1, CSF2, CSF3, CX3CL1,CXCL1, CXCL11, CXCL13, CXCL2, CXCL5, CXCL6, CXCL9, CXCR3, FGF2, GZMA, GZMB, HGF, IFNA1, IFNG, IL10, IL12A, IL12B, IL13, IL15, IL16, IL17A, IL18, IL1A, IL1B, IL2, IL20, IL21, IL22, IL23A, IL27, IL2RA, IL3, IL31, IL34, IL37, IL4, IL5, IL6, IL8, IL9, KITLG, LGALS3, LIF, LTA, MIF, NGF, PTX3, TNF, TNFRSF12A, TNFRSF1B, TNFRSF8, TNFSF10, TNFSF13, TNFSF13B, TREM1, TSLP, VEGFA, PPIB, HPRT1, GAPDH, GUSB | 1 plate | QGP-180-10080 |
3 plates | ||
| 10 plates | ||
QuantiGene Plex Mouse Immune Response Panel, 80-plex Targets: Il2ra, Il2, Il6, Ifng, Tnf, Il5, Il1a, Csf2, Il4, Il10, Il18, Vegfa, Il17a, Il1b, Il12a, Ccl2, Ccl7, Ccl11, Ccl5, Cxcl11, Il6ra, Ccl3, Ccl4, Il13, Il23a, Cxcr3, Il22, Il15, Il27, Il33, Ifna1, Il28a, Il31, Cxcl1, Cxcl2, Csf3, Il3, Lep, Tnfsf11, Csf1, Lif, Il9, Btc, Cxcl5, Il25, Il1rl1, Il19, Cd27, Kdr, Ccl19, Cxcl16, Il16, Ccl22, Ccl12, Gzmb, Ccl27a, Ccl24, Ccl17, Ccl25, Cxcl13, Il7r, Il7, Tslp, Tnfsf13b, Il12b, Il21, Gzma, Cd274, Ctla4, Cxcl9, Havcr2, Lag3, Ifna2, Ifnb1, Hgf, Tnfrsf12a, Ppib, Hprt, Gapdh, Gusb | 1 plate | |
3 plates | ||
10 plates | ||
QuantiGene Plex Human PanCancer Panel, 80-plex Targets: AGER, ARG1, AXL, BDNF, BTLA, CALR, CD27, CD274, CD276, CD28, CD36, CD47, CD48, CD80, CD96, CDH1, CSF3, CTLA4, CXCL8, DKK1, EGF, EPCAM, FGF2, GAPDH, GAS6, GPC1, GUSB, HAVCR2, HGF, HMGB1, HPRT1, HSP90AA1, HSPA4, HSPB2, HSPD1, ICOSLG, IDO1, IGF2, KITLG, LAG3, LGALS9, LIF, MBL2, MDK, MERTK, MICA, MICB, NCR3LG1, NECTIN2, NGF, NT5E, NTRK2, OLR1, PDCD1LG2, PECAM1, PGF, PLAUR, PPIB, PRF1, PVR, RAET1E, S100A8, S100A9, SERPINE1, SIGLEC7, SIGLEC9, SPARC, SPATA2, TIMD4, TNFRSF14, TNFRSF18, TNFRSF4, TNFRSF9, TRIM8, TYRO3, ULBP1, ULBP3, VEGFA, VEGFD, VSIR | 1 plate | |
3 plates | ||
10 plates | ||
QuantiGene Plex Mouse PanCancer Panel, 80-plex Targets: Ager, Arg1, Axl, Bdnf, Btla, Calr, Cd27, Cd274, Cd276, Cd28, Cd36, Cd47, Cd48, Cd80, Cd96, Cdh1, Csf3, Ctla4, Cxcl1, Dkk1, Egf, Epcam, Fgf2, Gapdh, Gas6, Gpc1, Gusb, Havcr2, Hgf, Hmgb1, Hprt, Hsp90Aa1, Hspa4, Hspb2, Hspd1, Icosl, Ido1, Igf2, Inhca , Kitl, Lag3, Lgals9, Lif, Mbl2, Mdk, Mertk, Mill2, Ncr1, Nectin2, Ngf, Nt5E, Ntrk2, Olr1, Pdcd1Lg2, Pecam1, Pgf, Plaur, Ppib, Prf1, Pvr, Raet1E, S100A8, S100A9, Serpine1, Siglece, Siglech, Sparc, Spata2, Timd4, Tnfrsf14 , Tnfrsf18, Tnfrsf4, Tnfrsf9, Trim8, Tyro3, Ulbp1, Ulbp3, Vegfa, Vegfd, Vsir | 1 plate | |
3 plates | ||
10 plates |
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