clec-87 Antibody
Description
Clarification of Terminology
The term "clec-87" does not correspond to any validated biological molecule or therapeutic antibody in peer-reviewed literature. A plausible explanation includes:
-
Typographical error: Confusion with CLEC-2 (a well-characterized platelet receptor) or other CLEC family members (e.g., CLEC-1, CLEC-3).
-
Misattribution: Potential conflation with unrelated CD antigens (e.g., CD85, CD32) or signaling molecules.
Overview of CLEC-2 and Its Antibodies
CLEC-2 is a type II transmembrane receptor expressed on platelets, immune cells, and endothelial cells. It binds podoplanin (a mucin-like glycoprotein) and activates platelet signaling via a hemITAM domain, contributing to thrombosis, tumor metastasis, and lymphangiogenesis . Antibodies targeting CLEC-2 or its ligands (e.g., podoplanin) are critical tools in research and therapeutic development.
Key CLEC-2-Targeting Antibodies and Their Mechanisms
4.1. Thrombosis and Hemostasis
-
INU1 (anti-CLEC-2) irreversibly removes CLEC-2 from platelets, mimicking the shedding mechanism observed with GPVI .
-
Deficiency models: CLEC-2 knockout mice exhibit prolonged bleeding times and resistance to arterial thrombosis .
4.2. Cancer Metastasis
-
Podoplanin-CLEC-2 interaction promotes platelet aggregation around tumor cells, facilitating metastasis .
-
Neutralizing antibodies (e.g., PG4D2, 2F7) block this interaction, reducing metastatic potential in preclinical models .
4.3. Immune Regulation
-
CLEC-2 signaling on dendritic cells enhances IL-10 production, modulating inflammation resolution .
-
BDCA2 (unrelated to CLEC-2) acts as an Fc receptor on plasmacytoid dendritic cells, regulating antibody-mediated immune responses .
Challenges and Future Directions
Product Specs
Composition: 50% Glycerol, 0.01M Phosphate Buffered Saline (PBS), pH 7.4
Q&A
Based on the analysis of available literature and technical resources, here is a structured FAQ addressing key research considerations for antibodies targeting CLEC-family receptors (adjusted from "CLEC-87" to align with validated CLEC-2/Dectin-1/AICL data in provided sources):
How to validate CLEC-family antibody specificity for flow cytometry?
Methodology:
-
Perform isotype control comparisons (e.g., Rat IgG2A for murine CLEC-2 ) and use knockout/transfected cell lines (e.g., EL4 cells transduced with HA-tagged CLEC-2 vs. vector controls ).
-
Combine with functional blocking assays (e.g., pre-incubation with recombinant CLEC-2-Fc protein to inhibit binding ).
Table 1: Validation Metrics
| Parameter | Example from CLEC-2 Study |
|---|---|
| Positive Control | Platelets (CD61+ cells) |
| Negative Control | Bone marrow neutrophils |
| Cross-reactivity | ≤10% with human Dectin-1 |
What are optimal conditions for detecting CLEC-2 in murine neutrophils?
Protocol:
-
Isolate peripheral blood neutrophils via Percoll gradient centrifugation (66%-78% interface ).
-
Stain within 2 hours post-collection to prevent receptor internalization.
-
Use anti-CD11b (5C6-FITC) and anti-Gr-1 for gating, with NaN₃ in buffers to inhibit internalization .
Key Finding:
CLEC-2 expression is 3–5× higher in peripheral blood neutrophils than bone marrow neutrophils (Fig. 1E-F ).
How to resolve contradictory CLEC-2 expression data across studies?
Analysis Framework:
-
Variable Sources:
-
Technical Factors:
Recommendation: Include strain-specific controls (BALB/c vs. C57BL/6 ) and stimulation timelines.
Designing functional assays for CLEC-2 signaling
Stepwise Approach:
-
Ligand Stimulation: Use rhodocytin (CLEC-2 agonist) at 50–100 nM .
-
Readouts:
-
Inhibition: Employ tyrosine-mutant chimeric receptors (Y7F in Dectin-1/CLEC-2 chimera ).
Data Conflict Example:
CLEC-2 activates Syk but lacks respiratory burst induction , unlike Dectin-1. Use phagocytosis assays (FITC-labeled beads ) to confirm functional divergence.
Combining CLEC antibodies with other therapies to avoid resistance
Strategy from SARS-CoV-2 Parallel :
-
Use non-competing antibody pairs (e.g., anti-CLEC-2 + anti-TLR2) to minimize escape mutants.
-
Preclinical testing:
-
In vitro: Expose to single vs. dual antibodies for 10–15 passages.
-
In vivo: Monitor variant emergence in hamster models over 14 days.
-
Table 2: Multi-Antibody Synergy
| Combination | Outcome | Source |
|---|---|---|
| REGEN-COV (dual) | 99% variant suppression | |
| CLEC-2 + Dectin-1 | Enhanced phagocytosis (1.8× ) |
