MSD V-PLEX · 111–131 Plex

Immune Checkpoints · T-Cell Effectors · TME Stroma

MSD Immuno-Oncology 111/131-Plex Panel Services

The most comprehensive soluble immune checkpoint and tumor microenvironment profiling panels on the MSD platform: 111 or 131 analytes covering PD-1, PD-L1, CTLA-4, LAG3, TIM-3, TIGIT, OX40, GITR, CD28, ICOS plus 60+ cytokines and chemokines, with the 131-Plex adding Granzyme B, Perforin, MMPs, and stromal interface markers — all from 25 µL of plasma.

Immunotherapy response is not determined by a single checkpoint — it emerges from the coordinated action of checkpoint signaling, T-cell effector function, cytokine-mediated immune polarization, chemokine-directed cell trafficking, and stromal barriers to immune infiltration. The 111-Plex captures immune checkpoints and cytokines; the 131-Plex adds the cytotoxic effector molecules (Granzyme A, Granzyme B, Perforin) that execute tumor cell killing, and the stromal markers (MMP-1/2/7/9, Galectin-9, E-Selectin, P-Selectin) that shape immune access to the tumor. Together they provide the most complete soluble biomarker profile available for Immuno-Oncologyresearch — from checkpoint engagement through T-cell activation to tumor cell killing and stromal remodeling.

Inquire About Immuno-Oncology Panels Compare 111 vs 131

Beyond Single Checkpoints — Systems-Level Immuno-Oncology Biomarker Profiling

The clinical success of checkpoint inhibitors proved that the immune system can be redirected against cancer. But the next generation of Immuno-Oncologyresearch demands moving beyond PD-L1 IHC as a single biomarker. Tumors evade immunity through multiple redundant checkpoint pathways simultaneously (PD-L1 + LAG3 + TIM-3 + TIGIT). T-cell exhaustion is a graded process — progressively losing IL-2 production, then TNF-α, then IFN-γ, then Granzyme B — that cannot be captured by any single marker. And the tumor stroma actively excludes T cells through MMP-mediated matrix remodeling, adhesion molecule modulation, and chemokine silencing — processes invisible to checkpoint-only assays. The 111/131-Plex panels address this complexity by measuring the full network in a single assay.

 Analyte Categories in the Immuno-Oncology Panels
Soluble immune checkpoints (15+): PD-1 (2 epitopes), PD-L1/B7-H1 (2 epitopes), PD-L2, CTLA-4, LAG3, HAVCR2/TIM-3, TIGIT, OX40/TNFRSF4, GITR/TNFRSF18, CD28, ICOS, CD27, HVEM/TNFRSF14, CD40L (soluble), ICOS-L/B7-H2, GITRL/TNFSF18 — covering inhibitory checkpoints, co-stimulatory receptors, and their ligands.
B-cell lineage & CAR-T targets: BAFF, BAFF-R/TNFRSF13C, BCMA/TNFRSF17, APRIL/TNFSF13, CD20 — B-cell biology, CAR-T target antigen monitoring (BCMA), and tertiary lymphoid structure research.
Cytokines & chemokines (60+): Full Th1/Th2/Th17/Treg cytokine panel plus chemokines directing immune cell trafficking (IP-10, MCP-1/2/4, MIP-1α/1β, MIG, I-TAC, RANTES) — capturing the cytokine milieu in which checkpoint blockade operates.
Cytotoxic effector molecules (131-Plex): Granzyme A, Granzyme B, Perforin — the three pore-forming and serine protease effectors of cytotoxic T lymphocytes and NK cells. Direct molecular readout of immune-mediated tumor cell killing.
Tumor-stroma interface (131-Plex): MMP-1, MMP-2, MMP-7, MMP-9 (Total), ProMMP-9, Galectin-9/GAL9, Nectin-4, E-Selectin, P-Selectin, Pentraxin 3, S100A12, RAGE/AGER — matrix remodeling enzymes, adhesion molecules, and damage-associated inflammatory mediators that shape TME architecture and immune access.
Additional immune regulators (131-Plex): LIGHT/TNFSF14, APRIL/TNFSF13, CD27, TLR1, gp130/sIL-6Rβ, CD276/B7-H3 — extending the immune signaling network coverage.

111-Plex vs. 131-Plex — Choosing the Right Configuration

Both configurations provide comprehensive checkpoint + cytokine coverage. The 131-Plex extends this with cytotoxic effector molecules and tumor-stroma interface markers for deeper TME characterization.

Analyte Category	111-Plex	131-Plex
Total Analytes	111	131
Soluble Immune Checkpoints	PD-1 (2 epitopes), PD-L1 (2 epitopes), PD-L2, CTLA-4, LAG3, TIM-3, TIGIT, OX40, GITR, CD28, ICOS, CD27, HVEM, CD40L	PD-1 (2 epitopes), PD-L1 (2 epitopes), PD-L2, CTLA-4, LAG3, TIM-3, TIGIT, OX40, GITR, CD28, ICOS, CD27, HVEM, CD40L, CD276/B7-H3
B-Cell & CAR-T Markers	BAFF, BAFF-R, BCMA, APRIL, CD20	BAFF, BAFF-R, BCMA, APRIL, CD20
Cytokines & Chemokines	60+ cytokines, chemokines, and growth factors	60+ cytokines, chemokines, and growth factors
Metabolic Hormones	Insulin, Leptin, C-Peptide, GLP-1, GIP, Glucagon, PYY, Ghrelin, FSH, LH, Proinsulin	Insulin, Leptin, C-Peptide, GLP-1, GIP, Glucagon, PYY, Ghrelin, FSH, LH, Proinsulin
Cytotoxic Effectors	Not included	Granzyme A, Granzyme B, Perforin
Matrix Remodeling	Not included	MMP-1, MMP-2, MMP-7, MMP-9 (Total), ProMMP-9
Adhesion & Stromal Interface	Not included	E-Selectin, P-Selectin, Nectin-4, Galectin-9, Pentraxin 3, S100A12, RAGE, TLR1, gp130, LIGHT, APRIL, GITRL, ICOS-L

The 111-Plex provides comprehensive immune receptor + cytokine coverage. The 131-Plex adds the functional readouts — are T cells actually killing tumor cells (Granzyme B, Perforin)? Is the stroma remodeled to permit immune access (MMPs, adhesion molecules)? Is there inflammatory amplification (Pentraxin 3, S100A12, RAGE)? The 20 additional analytes in the 131-Plex answer questions about immune function, not just immune receptor expression.

Immune Checkpoint Coverage — Detailed Analyte Listing

Inhibitory Checkpoints

PD-1 (epitope 1 & 2): dual-epitope assay distinguishes total PD-1 from drug-bound receptor — critical for differentiating PD-1 blockade occupancy from PD-1 expression changes. PD-L1/B7-H1 (epitope 1 & 2): dual-epitope for tumor- and immune-cell-derived sPD-L1. PD-L2: second PD-1 ligand. CTLA-4: the original checkpoint target. LAG3: co-expressed with PD-1 on exhausted T cells. HAVCR2/TIM-3: marks terminally exhausted T cells. TIGIT: competes with CD226 for CD155 binding on tumor cells.

Co-Stimulatory Receptors

CD28: the canonical T-cell co-stimulatory receptor — provides the second signal for T-cell activation. ICOS: inducible co-stimulator — amplifies T-cell responses and Tfh differentiation. OX40/TNFRSF4: promotes T-cell survival and memory formation; agonist antibodies in research studys. GITR/TNFRSF18: enhances effector T-cell function while suppressing Treg. CD27: co-stimulatory receptor promoting T-cell expansion. HVEM/TNFRSF14: bidirectional switch — inhibitory via BTLA or co-stimulatory via LIGHT. CD40L (soluble): the ligand for CD40 on APCs — bridges T-cell help to innate activation.

B-Cell & CAR-T Targets

BCMA/TNFRSF17: target of approved CAR-T therapies for multiple myeloma — soluble BCMA is a monitoring marker for CAR-T target antigen availability. BAFF & BAFF-R: B-cell survival factors. APRIL/TNFSF13: second BCMA ligand; also binds TACI. CD20: B-cell lineage marker; target of rituximab and bispecific antibodies. CD276/B7-H3 (131-Plex): emerging immunoregulatory B7 family member; CAR-T target in pediatric solid tumors.

Cytotoxic Effectors & Tumor-Stroma Interface — 131-Plex Exclusives

The 20 additional analytes in the 131-Plex that distinguish it from the 111-Plex — focused on immune effector function and TME architecture.

Cytotoxic Granules

Granzyme B — serine protease; initiates caspase-dependent and -independent apoptosis upon target cell entry via Perforin pores
Granzyme A — activates a parallel, caspase-independent cell death pathway
Perforin — forms pores in target cell membrane; prerequisite for granzyme entry
Elevated Granzyme B + Perforin during checkpoint therapy suggests T-cell activation; their absence despite checkpoint expression suggests functional exhaustion

Matrix Metalloproteinases

MMP-9 (Total) + ProMMP-9 — produced by TAMs and neutrophils; promotes angiogenesis and metastasis; ProMMP-9 reflects zymogen production rate
MMP-2 — degrades collagen IV in basement membrane; remodels ECM barriers to T-cell infiltration
MMP-1, MMP-7 — broad-spectrum matrix remodeling; MMP-7 also processes growth factors and cell surface proteins

Adhesion Molecules

E-Selectin, P-Selectin — endothelial adhesion molecules mediating leukocyte rolling and extravasation; gatekeepers of immune cell entry into tumor tissue
Nectin-4 — cell adhesion molecule upregulated in multiple solid tumors; target of enfortumab vedotin; shed form may serve as a pharmacodynamic marker

Inflammatory Mediators & DAMPs

Galectin-9 — binds TIM-3 on T cells, promoting exhaustion; also regulates macrophage polarization in the TME
Pentraxin 3 — soluble pattern recognition receptor; amplifies complement activation at tissue damage sites
S100A12 — neutrophil-derived DAMP; activates RAGE and TLR4 to propagate sterile inflammation
RAGE — receptor for S100 proteins, HMGB1, and AGEs; amplifies NF-κB-driven inflammatory signaling

Immuno-Oncology Panel Service Details

Service Package

Item	Full Service	Kit Only
Sample Processing	Included	— Self-run
111 or 131-Analyte Standard Curves	8-point, all analytes	Calibrators provided
QC Samples	Multi-level, duplicate	— User-prepared
Data Report	Organized by functional category	— Self-analysis
Raw ECL Data	Included	— Instrument-dependent

For Research Use Only. Not for diagnostic procedures.

How to Get Started

Select Configuration

Choose 111-Plex (checkpoint + cytokine) or 131-Plex (add effectors + stroma) based on whether T-cell cytotoxic function and stromal remodeling are central to your research.

Collect & Ship

EDTA plasma, processed within 1 hour, −80°C. Document treatment cycle, dosing, and concomitant medications for each timepoint.

Receive TME Profile

Receive your 111 or 131-analyte report organized by functional category — checkpoints, cytokines, effectors, stromal — with QC metrics.

MSD vs. Alternative Immuno-Oncology Biomarker Platforms

Feature	MSD (111/131-Plex)	Luminex xMAP
Max Plex	131 analytes in 1 well	Up to 50-plex
Soluble Checkpoint Coverage	15+ — most comprehensive single-panel checkpoint coverage	Limited — few validated checkpoint assays
Cytotoxic Effectors	Granzyme A/B, Perforin (131-Plex)	Limited
Sample Volume	25 µL	25–50 µL
Readout	Direct ECL — ~70 sec/plate	Flow cytometry — ~45–60 min/plate
Pre-Configured Immuno-Oncology Panel	Yes — V-PLEX with lot bridging	Custom premix required
Best Fit	Comprehensive Immuno-Oncologyprofiling with direct, rapid readout and documented checkpoint coverage	Budget-constrained screening with moderate plex

Immuno-Oncology Panel Frequently Asked Questions

What is the difference between the two PD-1 and PD-L1 epitope assays?

The dual-epitope assays for PD-1 and PD-L1 are designed to address a critical question in Immuno-Oncologyresearch: when you measure soluble PD-1 or PD-L1 in plasma from a research participant receiving checkpoint inhibitor immunotherapy, are you measuring the native protein or the drug-bound complex? In the 111/131-Plex, PD-1 epitope 1 detects total PD-1 regardless of drug binding; PD-1 epitope 2 may be masked when PD-1 is occupied by therapeutic antibody (nivolumab, pembrolizumab), allowing distinction between free and total PD-1. Similarly, PD-L1 epitope 1 and epitope 2 provide differential detection of free vs. drug-bound PD-L1 (atezolizumab, durvalumab, avelumab). This dual-epitope approach enables assessment of target engagement — has the drug saturated its target? — independent of target expression level. For research use only; panels use manufacturer-documented MSD reagents.

Can the 111-Plex data be compared with 131-Plex data if I switch configurations mid-study?

The 111 analytes common to both panels use identical antibody pairs, calibrators, and plate configurations. Data for these 111 analytes are directly comparable between the 111-Plex and 131-Plex. The 131-Plex simply adds 20 additional analytes on separate electrode spots. For longitudinal studies where you start with the 111-Plex and later upgrade to the 131-Plex, the core 111-analyte data is continuous. We recommend documenting the configuration used for each batch in your study metadata.

How should I handle the statistical challenge of 111–131 analytes measured simultaneously?

High-dimensional biomarker data requires appropriate statistical handling to avoid false positives. We recommend: (1) Pre-specify primary analytes based on your biological hypothesis — don't treat 111 independent tests as all primary endpoints; (2) Multiple testing correction (Benjamini-Hochberg) for exploratory analyses; (3) Composite signature approaches — combining multiple correlated analytes into composite scores reduces dimensionality and improves statistical power; (4) Discovery-validation split — use a subset of samples for discovery, then validate candidate biomarkers on an independent cohort. Our data reports include both raw and adjusted p-values. For advanced statistical analysis (PCA, hierarchical clustering, machine learning), our scientific team can provide consultation or refer you to bioinformatics collaborators.

Are these panels validated for monitoring immune-related adverse events (irAEs)?

No. These panels are for research use only and are not validated for clinical irAE monitoring or prediction. However, several panel analytes are of research interest in the context of irAEs: IL-6, TNF-α, and IL-1β are elevated in cytokine release syndrome; IL-17 and IL-23 have been associated with checkpoint inhibitor colitis; and soluble CD40L has been studied in the context of immune activation. The 111/131-Plex can be used to explore these associations in a research context, but results should not be used for clinical management of irAEs.

What sample collection metadata is essential for Immuno-Oncologybiomarker studies?

Critical metadata for each sample: (1) Treatment regimen and cycle number; (2) Days since last dose — drug levels influence soluble target measurements; (3) Concomitant medications — particularly corticosteroids (within 14 days), G-CSF, and other immunomodulatory drugs; (4) irAE status and grade — immune-related adverse events can produce acute-phase cytokine changes; (5) Baseline (pre-treatment) sample — essential for distinguishing treatment-induced changes from pre-existing inter-individual variability; (6) Best response (RECIST or iRECIST) — for retrospective biomarker-response correlation. We provide standardized IO-specific sample manifest forms for metadata capture. All services are research use only.

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