TFPI2 Antibody, FITC conjugated

What is TFPI2 and what are its primary biological functions?

TFPI2 (Tissue Factor Pathway Inhibitor 2), also known as Placental Protein 5 (PP5), is a secreted Kunitz-type serine proteinase inhibitor that plays a critical role in regulating plasmin-mediated matrix remodeling. The protein contains three Kunitz domains (KD1: residues 36-86, KD2: 96-149, and KD3: 158-208) separated by two linker regions, along with N-terminal acidic and C-terminal basic regions . TFPI2 primarily functions as a protease inhibitor that inhibits trypsin, plasmin, factor VIIa/tissue factor, and weakly inhibits factor Xa, but has no effect on thrombin . In cellular contexts, TFPI2 is abundantly expressed in non-invasive cells and the extracellular matrix but is often absent or undetectable in highly invasive tumor cells, suggesting its role as a tumor suppressor .

How is TFPI2 distributed within cells and tissues?

TFPI2 exhibits a complex cellular distribution pattern. While it is primarily secreted into the extracellular matrix (ECM) of various human tissues including liver, skeletal muscle, and pancreas, research has revealed that TFPI2 can also be found intracellularly . Both constitutively expressed TFPI2 and exogenously applied recombinant TFPI2 can be internalized and distributed throughout the cytoplasm and nucleus of cells . The nuclear localization is facilitated by a putative bipartite nuclear localization signal (NLS) in the C-terminal tail, and nuclear transport is mediated by the importin system . Immunofluorescence studies have shown that TFPI2 can be extensively expressed in cells, with predominant localization in the nucleus in certain cell types .

What structural domains are important for TFPI2 antibody epitope recognition?

The selection of appropriate antibodies for detecting TFPI2 depends on understanding its domain structure. TFPI2 contains an N-terminal acidic region, three Kunitz (K) domains, and a C-terminal basic region . Recombinantly produced TFPI2 antibodies may target various epitopes within this structure. Commercial antibodies like those from Abcam (ab86933, ab186747) are often raised against synthetic peptides within human TFPI2 . For specific experimental contexts, it's important to note that some commercial antibodies may target the N+KD1 regions (amino acids 1-95), while others may recognize the complete protein structure . When selecting a FITC-conjugated TFPI2 antibody, researchers should verify which epitope is targeted to ensure it aligns with their experimental objectives, especially if studying domain-specific protein interactions.

What are the optimal conditions for using FITC-conjugated TFPI2 antibodies in flow cytometry?

For optimal results in flow cytometry applications using FITC-conjugated anti-TFPI2 antibodies, the following protocol is recommended:

Sample Preparation:

• Harvest cells (1×10^6 cells per sample) and wash twice with cold PBS

• Fix cells with 4% paraformaldehyde for 10-15 minutes at room temperature

• Permeabilize with 0.1% Triton X-100 in PBS for 5-10 minutes if intracellular staining is required

Antibody Staining:

• Block non-specific binding with 1-3% BSA or normal serum for 30 minutes

• Incubate with FITC-conjugated anti-TFPI2 antibody at dilutions of 1:20-1:100 as recommended (e.g., BS-1144R-FITC)

• Incubate for 30-60 minutes at room temperature in the dark

• Wash three times with PBS to remove unbound antibody

• Resuspend cells in appropriate buffer for flow cytometric analysis

Controls and Analysis:

• Include appropriate isotype controls (FITC-conjugated rabbit IgG)

• Use unstained cells and single-stained controls for compensation if performing multicolor analysis

• Analyze using 488 nm laser excitation and appropriate emission filters for FITC detection (typically 530/30 nm)

This methodology ensures specific detection of TFPI2 while minimizing background fluorescence and non-specific binding.

How can I optimize immunofluorescence staining protocols for TFPI2 subcellular localization studies?

To effectively visualize TFPI2 subcellular localization using FITC-conjugated antibodies, consider this optimized protocol:

Cell Preparation:

• Culture cells on glass coverslips in appropriate media until 60-70% confluent

• Fix cells with 4% paraformaldehyde for 15 minutes at room temperature

• For nuclear localization studies, permeabilize with 0.2% Triton X-100 in PBS for 10 minutes

Staining Procedure:

• Block with 5% normal serum in PBS containing 0.1% Tween-20 (PBST) for 1 hour

• Incubate with FITC-conjugated anti-TFPI2 antibody at 1:50-1:200 dilution in blocking buffer overnight at 4°C

• Wash extensively (4-5 times) with PBST

• Counterstain nucleus with DAPI (1 μg/ml) for 5 minutes

• Mount using anti-fade mounting medium

For Co-localization Studies:

• When examining TFPI2 interaction with other proteins (e.g., myosin-9, actinin-4), perform double immunostaining by:

  • Using compatible secondary antibodies with distinct fluorophores

  • Following sequential staining for each primary antibody

  • Including appropriate controls for each antibody

Imaging Parameters:

• Use a confocal microscope with appropriate laser settings for FITC (excitation ~488 nm)

• Capture Z-stack images at 0.5-1 μm intervals to accurately assess nuclear localization

• Analyze colocalization using quantitative methods (e.g., Pearson's correlation coefficient)

This approach has successfully demonstrated that TFPI2 can be concentrated at the cell leading edge, colocalizing with myosin-9 (59%) and actinin-4 (52%) in cells overexpressing TFPI2 .

What controls should be included when performing co-immunoprecipitation experiments with TFPI2 antibodies?

For rigorous co-immunoprecipitation (Co-IP) experiments to study TFPI2 protein interactions, the following controls are essential:

Essential Controls:

• Input Control: Analyze 5-10% of the total cell lysate used for IP to confirm target protein expression

• Negative Control Antibody: Use isotype-matched IgG from the same species as the TFPI2 antibody

• Reciprocal IP: Perform reverse IP using antibodies against predicted interaction partners (e.g., myosin-9, actinin-4, SMURF2)

• Interaction Specificity Control: Include lysates from cells with TFPI2 knockdown or from cell lines that do not express TFPI2

• Bead-Only Control: Process lysate with beads alone (without antibody) to identify non-specific binding

Experimental Protocol Refinements:

• Pre-clear lysates with protein A/G beads to reduce non-specific binding

• Optimize lysis buffer conditions (standard recommendations: mammalian protein extraction reagent with protease inhibitors)

• For detecting transient interactions, consider using chemical crosslinking prior to cell lysis

• Wash stringently (4-5 times) with lysis buffer to remove non-specific interactions

Analysis and Validation:

• Confirm precipitation of TFPI2 by immunoblotting an aliquot of the precipitate with anti-TFPI2 antibody

• Validate novel interactions using alternative methods (e.g., proximity ligation assay, FRET)

• For nuclear interactions, perform nuclear fractionation prior to Co-IP

This approach has successfully identified TFPI2 interactions with proteins including importin-α , myosin-9, actinin-4 , and SMURF2 in various experimental models.

How can FITC-conjugated TFPI2 antibodies be employed to investigate its role in tumor suppression mechanisms?

FITC-conjugated TFPI2 antibodies offer powerful tools for investigating TFPI2's tumor suppression mechanisms through multiple advanced approaches:

Fluorescence-Based Cell Sorting and Analysis:

• Use flow cytometry to isolate TFPI2-expressing versus non-expressing populations from heterogeneous tumor samples

• Perform correlation analyses between TFPI2 expression levels and tumorigenic properties

• Monitor changes in TFPI2 expression during cancer progression or treatment response

High-Resolution Imaging of Signaling Dynamics:

• Employ time-lapse confocal microscopy to track TFPI2 translocation between cellular compartments following stimulation with TGF-β2 or other cytokines

• Visualize real-time interactions between TFPI2 and ERK signaling components to understand how TFPI2 regulates the EGFR/ERK1/2 pathway

• Use FRAP (Fluorescence Recovery After Photobleaching) to measure the mobility and binding dynamics of TFPI2 in different cellular compartments

Multiplex Imaging Approaches:

• Combine FITC-conjugated TFPI2 antibodies with markers for apoptosis (cleaved caspase-3/9) and proliferation (Ki-67) to simultaneously assess multiple cellular processes

• Implement multiplexed immunofluorescence to correlate TFPI2 expression with matrix metalloproteinases (especially MMP-2) and AP-2α to evaluate transcriptional regulation mechanisms

• Study co-localization with myosin-9 and actinin-4 at the leading edge of cells to understand TFPI2's role in regulating cell invasion

These methodologies have revealed that TFPI2 suppresses breast cancer cell proliferation by decreasing phosphorylation of ERK1/2 and inhibiting its nuclear translocation, while also repressing invasion through its interactions with cytoskeletal proteins.

What approaches can be used to study TFPI2 nuclear translocation and its effect on gene regulation?

Investigation of TFPI2 nuclear translocation and its gene regulatory functions requires sophisticated methodologies:

Nuclear Translocation Analysis:

• Subcellular Fractionation: Separate nuclear and cytoplasmic fractions followed by western blotting to quantify TFPI2 distribution

• Live-Cell Imaging: Use fluorescently tagged TFPI2 constructs alongside FITC-conjugated antibodies to track dynamic translocation in response to stimuli

• Nuclear Import Assays: Employ digitonin-permeabilized cell systems with recombinant TFPI2 to study importin-dependent nuclear transport mechanisms

Chromatin Association and Gene Regulation:

• ChIP-Seq Analysis: Perform chromatin immunoprecipitation followed by sequencing to identify TFPI2-associated genomic regions

• Transcription Factor Interaction Studies: Use sequential ChIP or Re-ChIP to determine if TFPI2 and AP-2α co-occupy the MMP-2 promoter region

• Promoter Activity Assays: Implement luciferase reporter constructs containing MMP-2 promoter regions to quantify the impact of TFPI2 on transcriptional activity

Molecular Mechanism Investigation:

• TFPI2 Domain Mapping: Generate TFPI2 deletion constructs lacking the nuclear localization signal (NLS) to confirm its requirement for nuclear entry and gene regulation

• Protein Complex Analysis: Employ proximity ligation assays to visualize and quantify TFPI2 interactions with transcription factors in the nuclear compartment

• Transcriptome Analysis: Compare RNA-seq profiles between wild-type, TFPI2-overexpressing, and NLS-deleted TFPI2 cells to identify globally regulated genes

Research using these approaches has demonstrated that TFPI2 can translocate to the nucleus and interact with transcription factor AP-2α, attenuating AP-2α binding to the MMP-2 promoter and consequently reducing MMP-2 transcription in breast cancer cells .

How can researchers differentiate between the extracellular and intracellular functions of TFPI2 using FITC-conjugated antibodies?

Distinguishing between extracellular and intracellular TFPI2 functions requires strategic experimental designs:

Differentiation Strategies:

• Non-permeabilized vs. Permeabilized Cell Staining:

  • Perform parallel immunofluorescence staining with FITC-conjugated TFPI2 antibodies on non-permeabilized cells (detecting only surface/extracellular TFPI2) and permeabilized cells (detecting total TFPI2)

  • Quantify the ratio of surface to total TFPI2 signal under different experimental conditions

• Targeted Inhibition Approaches:

  • Apply neutralizing antibodies against TFPI2 to the culture medium to specifically block extracellular functions

  • Compare with intracellular inhibition achieved through siRNA/shRNA knockdown or expression of dominant-negative TFPI2 constructs lacking specific domains

• Compartment-Specific Functional Assays:

  • Extracellular Function: Measure matrix degradation, plasmin activity, and MMP activation in conditioned media

  • Intracellular Function: Assess nuclear translocation of ERK1/2, AP-2α binding to the MMP-2 promoter, and gene expression changes

• Protein Engineering Approach:

  • Generate TFPI2 constructs with mutations in the signal peptide to prevent secretion while maintaining intracellular expression

  • Create fusion proteins with compartment-targeting signals to direct TFPI2 to specific subcellular locations

Analytical Methods:

• Use confocal microscopy with Z-stack imaging to precisely localize TFPI2 in cellular compartments

• Employ super-resolution microscopy techniques (STORM, PALM) for nanoscale visualization of TFPI2 distribution

• Combine with functional readouts such as invasion assays, proliferation measurements, and apoptosis detection

Research employing these strategies has revealed that TFPI2 functions distinctly in different cellular compartments: extracellularly inhibiting plasmin-mediated MMP activation , cytoplasmically interacting with cytoskeletal proteins like myosin-9 and actinin-4 , and nuclearly regulating gene expression through transcription factor interactions .

What are common problems encountered when using FITC-conjugated TFPI2 antibodies and how can they be resolved?

Researchers frequently encounter several challenges when working with FITC-conjugated TFPI2 antibodies:

Problem 1: Weak or Absent Signal

• Potential Causes:

  • Low endogenous TFPI2 expression (particularly in invasive cancer cell lines)

  • Suboptimal fixation masking epitopes

  • Antibody degradation due to improper storage or repeated freeze-thaw cycles

• Solutions:

  • Validate antibody on positive control cells known to express TFPI2

  • Optimize fixation protocols (test 4% PFA vs. methanol fixation)

  • Use signal amplification methods (tyramide signal amplification)

  • Store antibody in small aliquots at -20°C and protect from light

Problem 2: High Background or Non-specific Staining

• Potential Causes:

  • Insufficient blocking

  • Excessive antibody concentration

  • Autofluorescence from fixatives or cellular components

• Solutions:

  • Increase blocking time (2-3 hours) with 5% serum or BSA

  • Titrate antibody concentration (start with 1:50-1:200 dilutions)

  • Include 0.1-0.3% Triton X-100 in blocking buffer to reduce hydrophobic interactions

  • Use Sudan Black B (0.1-0.3%) to quench autofluorescence

  • Implement spectral unmixing during image acquisition

Problem 3: Discrepant Subcellular Localization Patterns

• Potential Causes:

  • Cell type-specific TFPI2 distribution

  • Epitope masking in specific compartments

  • Fixation artifacts affecting protein localization

• Solutions:

  • Compare multiple fixation methods

  • Validate localization with antibodies targeting different TFPI2 epitopes

  • Confirm with complementary approaches (e.g., TFPI2-GFP fusion proteins)

  • Consider cell-specific factors that might influence TFPI2 trafficking

Problem 4: Photobleaching During Imaging

• Potential Causes:

  • Extended exposure to excitation light

  • Suboptimal mounting medium

• Solutions:

  • Use anti-fade mounting media containing radical scavengers

  • Minimize exposure time and laser power during acquisition

  • Consider alternative conjugates with greater photostability than FITC

Each of these solutions has been validated in research contexts examining TFPI2 localization and function in various cell types.

How do you interpret conflicting data between TFPI2 antibody-based detection methods and gene expression analysis?

Discrepancies between protein detection and gene expression data for TFPI2 require systematic investigation:

Common Discrepancy Scenarios and Analytical Approaches:

• High mRNA Expression with Low/Undetectable Protein:

  • Potential Mechanisms:

    • Post-transcriptional regulation (miRNAs, RNA-binding proteins)

    • Rapid protein degradation

    • Secretion and matrix binding reducing cellular detection

  • Verification Methods:

    • Assess extracellular matrix for TFPI2 deposition

    • Use proteasome inhibitors to block potential degradation

    • Analyze conditioned media for secreted TFPI2

    • Measure protein half-life with cycloheximide chase assays

• Detectable Protein with Low mRNA Expression:

  • Potential Mechanisms:

    • High protein stability/slow turnover

    • Internalization of exogenous TFPI2 from culture medium

    • Antibody cross-reactivity with related proteins

  • Verification Methods:

    • Validate antibody specificity using TFPI2 knockout/knockdown controls

    • Perform pulse-chase experiments to determine protein stability

    • Test for TFPI2 internalization using recombinant protein

• Subcellular Localization Inconsistencies:

  • Potential Mechanisms:

    • Cell type-specific trafficking patterns

    • Condition-dependent nuclear translocation

    • Epitope masking in certain compartments

  • Verification Methods:

    • Compare multiple antibodies targeting different TFPI2 domains

    • Validate with fractionation followed by western blotting

    • Use GFP-tagged TFPI2 constructs to confirm localization patterns

Integrated Analysis Framework:

• Temporal Considerations: Analyze whether mRNA changes precede protein changes

• Quantitative Correlation: Determine if there's a non-linear relationship between mRNA and protein levels

• Context-Specific Regulation: Evaluate if discrepancies occur under specific conditions (e.g., TGF-β2 stimulation)

• Isoform Analysis: Investigate if specific TFPI2 isoforms or post-translationally modified forms are differentially detected

Research has shown that TFPI2 undergoes complex regulation, including internalization and nuclear translocation , which can contribute to these discrepancies and must be considered during data interpretation.

What are the most effective strategies for validating the specificity of FITC-conjugated TFPI2 antibodies?

Rigorous validation of FITC-conjugated TFPI2 antibodies requires a multi-faceted approach:

Essential Validation Strategies:

• Genetic Validation Approaches:

  • TFPI2 Knockdown/Knockout Controls: Confirm signal reduction in TFPI2-silenced cells using siRNA, shRNA, or CRISPR-Cas9

  • Overexpression Systems: Verify increased signal in cells transfected with TFPI2 expression constructs

  • Rescue Experiments: Restore TFPI2 expression in knockout cells and confirm antibody signal recovery

• Biochemical Validation:

  • Peptide Competition Assays: Pre-incubate antibody with purified TFPI2 or immunizing peptide before staining to block specific binding

  • Western Blot Correlation: Confirm that immunofluorescence patterns match band patterns observed in western blots (expected size: 27 kDa; observed size: 25 kDa)

  • Multiple Antibody Comparison: Test several antibodies targeting different TFPI2 epitopes to confirm consistent staining patterns

• Biological Context Validation:

  • Tissue/Cell Expression Patterns: Verify that staining matches known TFPI2 expression patterns (e.g., high in non-invasive cells, low in invasive cancer cell lines)

  • Functional Correlation: Confirm that TFPI2 staining correlates with expected biological activities (e.g., reduced ERK phosphorylation, decreased invasion)

  • Induction Experiments: Demonstrate appropriate signal changes following treatments known to alter TFPI2 expression

• Technical Controls:

  • Isotype Controls: Use isotype-matched FITC-conjugated IgG from the same species

  • Absorption Controls: Pre-absorb antibody with recombinant TFPI2 protein

  • Secondary-Only Controls: For indirect immunofluorescence methods

Advanced Validation for Specialized Applications:

• For nuclear localization studies: Validate with subcellular fractionation followed by western blotting

• For co-localization experiments: Perform proximity ligation assays to confirm protein-protein interactions identified by co-immunofluorescence

• For internalization studies: Track antibody uptake in real-time using live-cell imaging

Researchers have successfully employed these validation approaches to confirm TFPI2 antibody specificity in studies examining its role in tumor suppression and complex cellular trafficking patterns .

How are FITC-conjugated TFPI2 antibodies being used to investigate its role in cancer progression and metastasis?

FITC-conjugated TFPI2 antibodies have become instrumental in elucidating TFPI2's functions in cancer:

Current Applications in Cancer Research:

• Tumor Microenvironment Interactions:

  • Multichannel Fluorescence Imaging: Visualizing TFPI2 expression patterns relative to extracellular matrix components and invasive tumor fronts

  • 3D Spheroid Invasion Models: Tracking TFPI2 dynamics during invasion into surrounding matrices

  • Co-culture Systems: Examining TFPI2 exchange between cancer cells and stromal components

• Metastatic Process Investigation:

  • Circulating Tumor Cell Analysis: Quantifying TFPI2 expression in CTCs as a potential biomarker for metastatic potential

  • Intravasation/Extravasation Models: Studying TFPI2's role during critical steps of the metastatic cascade

  • Metastatic Niche Formation: Evaluating how TFPI2 influences cancer cell colonization at distant sites

• Therapeutic Response Monitoring:

  • Treatment-Induced Changes: Measuring alterations in TFPI2 expression and localization following chemotherapy, targeted therapy, or immunotherapy

  • Resistant Phenotype Characterization: Comparing TFPI2 patterns between treatment-responsive and resistant cells

  • Combination Therapy Optimization: Assessing whether modulating TFPI2 can enhance response to standard therapies

Insights from Recent Research:

• TFPI2 overexpression decreases phosphorylation of EGFR/ERK1/2 and inhibits ERK1/2 nuclear translocation, reducing cell proliferation in breast cancer models

• Interaction of TFPI2 with myosin-9 and actinin-4 inhibits cell migration and invasion potential

• Nuclear localization of TFPI2 contributes to inhibition of MMP-2 transcription by interfering with AP-2α binding to the MMP-2 promoter

• Restoration of TFPI2 in glioblastoma cells triggers both intrinsic and extrinsic caspase-mediated apoptotic pathways

These applications have revealed TFPI2 as a multifunctional tumor suppressor that acts through diverse mechanisms affecting proliferation, invasion, and survival pathways in cancer cells.

What novel techniques are being developed to study TFPI2 interactions with signaling molecules using fluorescence-based approaches?

Cutting-edge fluorescence techniques are advancing our understanding of TFPI2's roles in signaling networks:

Emerging Methodologies:

• Advanced Live-Cell Imaging Approaches:

  • FRET-Based Biosensors: Developing TFPI2-based Förster Resonance Energy Transfer sensors to monitor real-time interactions with binding partners such as SMURF2, myosin-9, or actinin-4

  • Optogenetic TFPI2 Regulation: Using light-controlled systems to manipulate TFPI2 localization or activity with spatiotemporal precision

  • Super-Resolution Live Imaging: Applying techniques like lattice light-sheet microscopy to track TFPI2 dynamics at nanoscale resolution

• Single-Molecule Approaches:

  • Single-Molecule Tracking: Following individual TFPI2 molecules labeled with photo-convertible fluorophores to map mobility patterns and binding kinetics

  • Single-Cell Protein-Protein Interaction Analysis: Using techniques like fluorescence correlation spectroscopy (FCS) to measure TFPI2 interaction affinities in living cells

  • Expansion Microscopy: Physically expanding fixed samples to achieve super-resolution imaging of TFPI2 and its interaction partners

• Multiparametric Systems-Level Analysis:

  • Multiplexed Imaging: Combining FITC-conjugated TFPI2 antibodies with panels of signaling markers for simultaneous visualization of multiple pathway components

  • Mass Cytometry Imaging: Adapting metal-tagged antibodies against TFPI2 and signaling molecules for highly multiplexed tissue analysis

  • Spatial Transcriptomics Integration: Correlating TFPI2 protein localization with transcriptional outputs at single-cell resolution

Specific Applications to TFPI2 Signaling Studies:

• Investigating dynamic changes in TFPI2-SMURF2 interactions following TGF-β2 stimulation using proximity ligation assays

• Tracking TFPI2's effect on ERK1/2 phosphorylation and nuclear translocation kinetics using live-cell reporters

• Visualizing the assembly and disassembly of TFPI2 complexes with cytoskeletal proteins at the leading edge of migrating cells

These innovative approaches are uncovering how TFPI2 functions as a critical regulator at the intersection of multiple signaling pathways, including TGF-β/Smad, ERK, and cytoskeletal remodeling networks.

How might TFPI2 antibodies contribute to therapeutic developments targeting cancer and fibrotic diseases?

TFPI2 antibodies hold significant potential for translational and therapeutic applications:

Therapeutic Strategy Development:

• Diagnostic and Prognostic Applications:

  • Precision Medicine Biomarkers: Using FITC-conjugated TFPI2 antibodies to stratify patients based on TFPI2 expression profiles

  • Circulating Tumor Cell (CTC) Analysis: Developing TFPI2-based CTC detection systems for early metastasis detection

  • Treatment Response Monitoring: Tracking changes in TFPI2 expression patterns during therapy

• Targeted Therapy Design:

  • Domain-Specific Inhibitors: Developing therapeutic agents targeting specific TFPI2 interactions based on antibody epitope mapping data

  • Protein-Protein Interaction Modulators: Designing small molecules that mimic TFPI2's interaction with critical partners like SMURF2

  • Nuclear Translocation Enhancers: Creating compounds that promote TFPI2 nuclear localization to enhance its tumor-suppressive transcriptional regulation

• TFPI2 Restoration Strategies:

  • Epigenetic Modifiers: Developing drugs that reverse TFPI2 promoter methylation in cancers

  • mRNA Stabilization: Targeting microRNAs that regulate TFPI2 expression

  • Protein Delivery Systems: Engineering nanoparticles for targeted delivery of recombinant TFPI2 to tumor sites

Disease Applications Beyond Cancer:

• Diabetic Nephropathy: Targeting TFPI2's role in endothelial-mesenchymal transition (EndMT) and fibrosis

• Inflammatory Diseases: Exploring TFPI2's functions in regulating tissue factor pathways and coagulation

• Neurodegenerative Conditions: Investigating TFPI2's potential neuroprotective effects through matrix remodeling regulation

Methodological Considerations:

• Antibody-based imaging could guide surgical resection margins by visualizing TFPI2 expression boundaries

• Developing antibody-drug conjugates targeting cells with aberrant TFPI2 expression patterns

• Creating bispecific antibodies linking TFPI2 status to immune effector recruitment