LUMINEX MULTIPLEX ASSAY SERVICE

Mouse Immuno-Oncology Checkpoint Panel (7/11-Plex)

Simultaneous quantification of key mouse immune checkpoint proteins — including the PD-1/PD-L1 axis, CTLA-4, co-stimulatory ligands, and B7 family members — in a single well using Luminex xMAP technology. Designed for preclinical cancer immunotherapy research, syngeneic tumor model studies, and immuno-oncology drug development.

7/11 TargetsMouse25 μL Samplepg/mL Sensitivity
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7/11-Plex Immuno-Oncology
Brown University
Harvard University
Imperial College London
University of Florida
Tulane University
Abata Therapeutics
AlzeCure Pharma

The adaptive immune system maintains a delicate balance between T cell activation and inhibition through a network of co-stimulatory and co-inhibitory receptors and their ligands, collectively known as immune checkpoints. In cancer, tumor cells and the tumor microenvironment exploit these checkpoints — particularly the PD-1/PD-L1/PD-L2 axis and CTLA-4 — to suppress anti-tumor T cell responses. Immune checkpoint inhibitors (anti-PD-1, anti-PD-L1, anti-CTLA-4 antibodies) have revolutionized cancer therapy, and syngeneic mouse tumor models are the primary preclinical platform for evaluating these agents.

Creative Proteomics offers the Mouse Immuno-Oncology Checkpoint Panel based on the Luminex xMAP platform for simultaneous quantification of key mouse immune checkpoint proteins in a single well. The panel covers the PD-1/PD-L1/PD-L2 axis (the dominant inhibitory checkpoint), CTLA-4 + CD80 (the co-stimulatory/co-inhibitory pair), and B7-H3 + CD137L (emerging checkpoint and co-stimulatory targets). The 7-plex core configuration can be expanded to 11-plex with additional targets. Validated for serum, plasma, and cell culture supernatants, the panel is compatible with MAGPIX, Luminex 200, and FLEXMAP 3D systems.

The panel captures soluble isoforms of checkpoint proteins shed or secreted into the circulation and tumor microenvironment, enabling non-invasive monitoring of checkpoint pathway activity in preclinical mouse models from a single 25 μL sample per time point.

Panel Specifications
TechnologyLuminex xMAP
Panel Size7-plex (core), expandable to 11-plex
SpeciesMouse
Sample TypesSerum, EDTA/Heparin Plasma, CCS
Sample Volume25 μL (serum/plasma), 50 μL (CCS)
SensitivitySub-pg/mL to pg/mL
Assay Time~4 hours

Complete Analyte List — Mouse Immune Checkpoint Targets

The Mouse Immuno-Oncology Checkpoint Panel measures soluble checkpoint proteins across three functional categories: the PD-1 axis, co-stimulatory/co-inhibitory receptors, and B7 family ligands.

Target Alternative Name Functional Category Biological Function in Immuno-Oncology
PD-1 CD279, PDCD1 Co-Inhibitory Receptor Dominant T cell exhaustion marker; upregulated on chronically stimulated tumor-infiltrating T cells; engagement by PD-L1/PD-L2 suppresses TCR signaling and effector function; primary target of checkpoint inhibitor immunotherapy (anti-PD-1)
PD-L1 CD274, B7-H1 Co-Inhibitory Ligand Expressed on tumor cells, tumor-associated macrophages, and myeloid-derived suppressor cells; IFN-γ-inducible; directly suppresses PD-1+ T cells in the tumor microenvironment; predictive biomarker for anti-PD-1/PD-L1 therapy response
PD-L2 CD273, B7-DC Co-Inhibitory Ligand Second ligand for PD-1; predominantly expressed on antigen-presenting cells; binds PD-1 with 2–6 fold higher affinity than PD-L1; may mediate PD-1 signaling when PD-L1 is low or blocked
CTLA-4 CD152 Co-Inhibitory Receptor High-affinity competitor for CD80/CD86; outcompetes CD28 for ligand binding, raising the threshold for T cell activation; constitutively expressed on Tregs; target of ipilimumab-class checkpoint inhibitors; soluble CTLA-4 isoform generated by alternative splicing
CD80 B7-1 Co-Stimulatory/Co-Inhibitory Ligand Shared ligand for CD28 (co-stimulatory) and CTLA-4 (co-inhibitory); expressed on activated APCs and some tumor cells; provides positive signal through CD28 in priming phase; provides negative signal through CTLA-4 in effector phase
B7-H3 CD276 Co-Inhibitory/Immune Evasion Broadly expressed on tumor cells and tumor vasculature; inhibits T cell proliferation and effector function; associated with poor prognosis across multiple cancer types; emerging therapeutic target (anti-B7-H3 ADCs and bispecific antibodies in clinical development)
CD137L 4-1BBL, TNFSF9 Co-Stimulatory Ligand Ligand for CD137 (4-1BB), a potent T and NK cell co-stimulatory receptor; expressed on activated APCs; engagement promotes T cell survival, memory formation, and anti-tumor cytotoxicity; target of agonistic anti-4-1BB antibodies in immuno-oncology
Why measure soluble checkpoint proteins? Membrane-bound immune checkpoints (PD-L1 on tumor cells, CTLA-4 on T cells) are assessed by immunohistochemistry or flow cytometry, which require tissue biopsy. Soluble checkpoint isoforms (sPD-L1, sPD-1, sCTLA-4, sCD80, sB7-H3) are shed or secreted into serum and tumor interstitial fluid, enabling non-invasive, serial monitoring of checkpoint pathway activity from a blood draw. In mouse models where serial tumor biopsies are impractical due to animal welfare constraints and limited tissue mass, soluble checkpoint measurement is particularly valuable for longitudinal pharmacodynamic assessment.

Don't see the checkpoint target you need? This 7-plex panel can be expanded to 11-plex with additional validated targets from the mouse immuno-oncology platform, including co-stimulatory receptors and additional B7 family ligands. Contact us with your target list for a custom panel quote.

Technical Specifications

Validated performance parameters for the Mouse Immuno-Oncology Checkpoint Panel.

Platform and Assay
PlatformLuminex xMAP (MAGPIX / Luminex 200 / FLEXMAP 3D)
Panel Configurations7-plex (core), expandable to 11-plex
SpeciesMouse
Sample TypesSerum, EDTA/Heparin Plasma, CCS
Sample Volume25 μL (serum/plasma), 50 μL (CCS)
Assay Time~4 hours
Performance Metrics
SensitivitySub-pg/mL to pg/mL (varies by analyte)
Dynamic Range3–4 logs
Intra-Assay CV<10%
Inter-Assay CV<15%
Spike Recovery80–120%
Standard Curve5PL fit, R² >0.99

Soluble vs. Membrane-Bound Checkpoint Proteins

Immune checkpoint proteins exist in two forms: membrane-bound (requiring biopsy) and soluble (detectable in serum/plasma). Understanding the distinction is critical for interpreting multiplex data in preclinical mouse models.

Parameter Soluble Checkpoint (This Panel) Membrane-Bound Checkpoint (IHC / Flow)
Measurement Serum/plasma concentration via Luminex Tissue expression via IHC or flow cytometry
Sample Requirement 25 μL blood Tumor biopsy or tissue section
Serial Monitoring Yes — non-invasive, repeatable No — terminal procedure in mice
PD-L1 Detection sPD-L1 shed from tumor and immune cells Membrane PD-L1 on tumor cells (IHC H-score)
CTLA-4 Detection sCTLA-4 alternatively spliced isoform Intracellular CTLA-4 in Tregs (flow cytometry)
Best Use Case Longitudinal pharmacodynamic monitoring; treatment response assessment Baseline characterization; tumor microenvironment spatial context

In syngeneic mouse models (MC38, B16-F10, CT26, 4T1), soluble checkpoint protein levels change dynamically during tumor growth and in response to checkpoint inhibitor therapy. Serum sPD-L1 typically rises with tumor burden and declines following effective anti-tumor treatment. The 7-plex panel enables tracking of these dynamic changes from serial blood samples without sacrificing animals at each time point.

Sample Requirements for Mouse Immuno-Oncology Checkpoint Assays

Mouse blood volume is limited (~1.5–2 mL total for a 25 g mouse). The 25 μL requirement for this panel enables serial sampling in longitudinal studies.

Sample Type Volume Requirement
Serum 25 μL Collect via submandibular or retro-orbital bleed; allow clotting for 30 min at room temperature; centrifuge at 1,500g for 10 min. Serial sampling: 50–75 μL whole blood yields 25 μL serum. Terminal collection: cardiac puncture.
EDTA/Heparin Plasma 25 μL Centrifuge within 30 min at 2,500g for 15 min; EDTA preferred over heparin for checkpoint proteins
Tumor Interstitial Fluid 50 μL Collect tumor, rinse in PBS, incubate in buffer at 4°C for 1 hour, centrifuge at 14,000g for 15 min; provides localized checkpoint concentration data complementary to systemic serum levels
Cell Culture Supernatant 50 μL Centrifuge at 10,000g for 10 min; for tumor cell line conditioned media or splenocyte/T cell co-culture experiments
Minimum Project Size — One 96-well plate; smaller batches accepted with surcharge
Sample Storage — -80°C; avoid repeated freeze-thaw cycles
Shipping — Dry ice; samples must remain frozen throughout transit

How the Mouse Immuno-Oncology Checkpoint Panel Works

The panel measures soluble checkpoint proteins across three functional axes, providing a comprehensive view of the immuno-oncology signaling landscape in mouse models.

PD-1

PD-1/PD-L1/PD-L2 Axis

PD-1, PD-L1, PD-L2 — the dominant T cell exhaustion pathway targeted by the most widely used checkpoint inhibitors (anti-PD-1, anti-PD-L1). In syngeneic models, serum sPD-L1 typically rises with tumor burden and declines with effective anti-PD-1 therapy. 25 μL per well.

Therapeutic targets: anti-PD-1, anti-PD-L1
CTLA-4

CTLA-4 / CD80 / CD86 Pathway

CTLA-4, CD80 — CTLA-4 is the high-affinity competitor for CD80/CD86 that raises the T cell activation threshold. sCTLA-4 is an alternatively spliced isoform with immunomodulatory activity. CD80 is the shared ligand providing co-stimulation (via CD28) or co-inhibition (via CTLA-4). 25 μL per well.

Therapeutic targets: anti-CTLA-4
Emerging

Emerging Checkpoint Targets

B7-H3, CD137L — B7-H3 is a broadly expressed tumor immune evasion molecule. CD137L engages CD137 (4-1BB) to provide co-stimulatory signals critical for T cell survival and anti-tumor memory. Both are active targets in clinical-stage immuno-oncology programs. 25 μL per well.

Emerging targets: anti-B7-H3 ADCs, anti-4-1BB agonists
Synergy with Human Immune Checkpoint 37-Plex: The mouse panel is designed as the preclinical counterpart to our Human Immune Checkpoint 37-Plex Panel. In a typical drug development workflow, the mouse 7/11-plex is used for in vivo PD studies in syngeneic models, while the human 37-plex is used for patient sample analysis in clinical trials. The same Luminex platform, the same sample volume, and the same data format ensure seamless translation from mouse pharmacodynamics to human biomarker analysis.

Mouse Immuno-Oncology Checkpoint Panel Research Applications

The panel supports preclinical cancer immunotherapy research, checkpoint inhibitor pharmacodynamics, and syngeneic tumor model studies.

Syngeneic Tumor Model Pharmacodynamics

Monitor soluble checkpoint protein changes in serum during tumor growth and checkpoint inhibitor treatment in standard syngeneic models (MC38, B16-F10, CT26, 4T1). sPD-L1 levels reflect tumor burden and response to therapy. Serial measurement from 25 μL blood samples enables dense time-course PK/PD studies without terminal sampling.

Checkpoint Inhibitor Efficacy Studies

Quantify target engagement and pharmacodynamic response for anti-PD-1, anti-PD-L1, anti-CTLA-4, and combination therapies. Rising sPD-L1 during treatment may indicate adaptive immune resistance via IFN-γ-driven PD-L1 upregulation — a known mechanism of acquired resistance detectable by the panel.

Novel Immunotherapy Target Validation

Evaluate next-generation immuno-oncology agents targeting B7-H3, 4-1BB (CD137), and other emerging checkpoint pathways. The B7-H3 and CD137L measurements enable target expression monitoring, while the PD-1 axis markers provide context for combination therapy studies.

Tumor Microenvironment Characterization

Compare checkpoint protein concentrations in serum vs. tumor interstitial fluid to distinguish systemic from local (tumor microenvironment) checkpoint pathway activity. This paired analysis reveals whether soluble checkpoint elevation reflects tumor-localized immune evasion or systemic immune modulation.

Combination Immunotherapy Assessment

Evaluate checkpoint inhibitor combinations (anti-PD-1 + anti-CTLA-4, anti-PD-L1 + anti-4-1BB) by measuring the full checkpoint ligand/receptor panel. The panel detects compensatory upregulation (e.g., PD-L2 increase during anti-PD-L1 monotherapy) that may inform combination partner selection.

Mouse-to-Human Translational Biomarker Studies

Use the mouse panel for preclinical biomarker discovery, then translate findings to clinical studies using our Human Immune Checkpoint 37-Plex Panel. The shared platform, overlapping targets, and compatible data formats enable direct cross-species biomarker comparison.

Deliverables and Quality Metrics

Every Luminex mouse immuno-oncology checkpoint assay includes a comprehensive data package with full quality control documentation.

Data Package
  • Raw fluorescence intensities (.csv)
  • Calculated concentrations (pg/mL) for all checkpoint targets
  • 5PL standard curves for each analyte (R² >0.99)
  • Full QC report (.xlsx format)
Quality Control
  • Standard curve: 7-point dilution series, 5PL fit, R² >0.99
  • Intra-assay CV <10%
  • Inter-assay CV <15%
  • Spike recovery: 80–120%
Assay Performance
  • Duplicate sample measurements for all samples
  • Method summary with reagent lot numbers
  • LLOQ reported per analyte
  • Platform: Luminex xMAP, compatible with MAGPIX, Luminex 200, FLEXMAP 3D

Related Panels

Explore other Luminex panels available for immuno-oncology research across species.

Human Immune Checkpoint 37-Plex Panel
  • Targets: PD-1, PD-L1, CTLA-4, TIM-3, LAG-3, B7-H3, CD28, OX40, 4-1BB
  • Platform: Luminex
  • Plex: 37-Plex
View Details
Human Cell Therapy Cytokine Panel
  • Targets: IL-6, IFN-γ, GM-CSF, IL-10, IL-2, IP-10
  • Platform: Luminex
  • Plex: 20/25-Plex
View Details
Mouse Tumor Biomarker Panel
  • Targets: Tumor biomarker cytokines
  • Platform: Luminex
  • Plex: Multiplex
View Details
Mouse Cytokine Panel
  • Targets: Comprehensive mouse cytokine profiling
  • Platform: Luminex
  • Plex: Multiplex
View Details
Mouse Immune Monitoring Panel
  • Targets: Cytokines, chemokines, immune markers
  • Platform: Luminex
  • Plex: Multiplex
View Details
Mouse Oncology Panel
  • Targets: Oncology biomarkers
  • Platform: Luminex
  • Plex: Multiplex
View Details
← All Mouse PanelsBrowse mouse species overview

Frequently Asked Questions About Mouse Immuno-Oncology Checkpoint Panel

Common questions about our mouse immuno-oncology checkpoint Luminex multiplex panel service.

How does this mouse panel relate to the Human Immune Checkpoint 37-Plex?
The Mouse Immuno-Oncology Checkpoint Panel (7/11-plex) is designed as the preclinical counterpart to the Human Immune Checkpoint 37-Plex. It includes the core targets most relevant to syngeneic mouse model studies: the PD-1/PD-L1/PD-L2 axis, CTLA-4/CD80, and emerging targets B7-H3 and CD137L. The human 37-plex provides broader coverage with 37 targets, including many (TIM-3, LAG-3, OX40, GITR) that have limited or different biology in mice. For translational studies, use the mouse panel for in vivo pharmacodynamics and the human panel for clinical biomarker analysis.
Why soluble checkpoint proteins instead of membrane-bound IHC or flow?
IHC and flow cytometry require tumor tissue, which in mice means terminal sacrifice at each time point. To assess checkpoint pathway dynamics over a 3-week treatment course at 3 time points, IHC would require 3 cohorts of n=10 mice (30 total). Soluble checkpoint measurement from 25 μL serum samples enables serial bleeding (submandibular or tail vein) from the same 10 mice at all 3 time points — reducing animal numbers by 67% and enabling within-subject longitudinal analysis that is statistically more powerful than cross-sectional comparisons.
What syngeneic tumor models are compatible with this panel?
The panel is validated for mouse serum and plasma from all standard syngeneic tumor models, including MC38 (colon adenocarcinoma, C57BL/6), B16-F10 (melanoma, C57BL/6), CT26 (colon carcinoma, BALB/c), 4T1 (mammary carcinoma, BALB/c), LLC (Lewis lung carcinoma, C57BL/6), and Pan02 (pancreatic adenocarcinoma, C57BL/6). The panel is species-specific (mouse) but strain-independent; it works across C57BL/6, BALB/c, FVB, and other common inbred strains.
How much blood can I safely collect from a mouse for serial checkpoint monitoring?
The panel requires 25 μL of serum, which translates to ~50–75 μL of whole blood per time point. Under IACUC guidelines, up to 10% of total blood volume (~150–200 μL for a 25 g mouse) can be safely collected every 2 weeks, or 1% (~15–20 μL) daily. For a typical 21-day tumor growth study with weekly sampling (Days 0, 7, 14, 21), the total blood collection (<300 μL over 3 weeks) is well within approved limits. Submandibular or tail vein collection with 25G needles is recommended.
Can this panel detect treatment-induced checkpoint upregulation?
Yes. A well-characterized response to anti-PD-1 or anti-PD-L1 therapy is IFN-γ-driven PD-L1 upregulation (adaptive immune resistance). This manifests as rising serum sPD-L1 during treatment despite tumor control. The 7-plex panel detects this pharmacodynamic signal because it measures sPD-L1 directly. Similarly, anti-CTLA-4 therapy may alter CD80 levels on APCs, and the panel quantifies sCD80 in parallel. These measurements distinguish true pharmacodynamic response from simple tumor burden correlation.
Can this panel be expanded beyond 7 targets?
Yes. The 7-plex core configuration can be expanded to 11-plex with additional targets from the mouse immuno-oncology portfolio, including co-stimulatory receptors and additional B7 family ligands. Contact us with your specific targets of interest for a custom panel configuration. Note: the panel is pre-configured for optimal cross-reactivity and cannot be arbitrarily combined with arbitrary simplex assays; additions are limited to validated complementary targets within the same platform family.

Interested in Mouse Immuno-Oncology Profiling?

Contact us to discuss your syngeneic tumor model study requirements, checkpoint panel configuration (7-plex or 11-plex), and serial sampling protocols. We respond within 24 hours.

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