unc-71 Antibody
Description
UNC-71 Protein in C. elegans
UNC-71 is a disintegrin and metalloprotease (ADAM) protein encoded by the unc-71 gene in Caenorhabditis elegans. Key findings include:
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Function: Regulates motor axon guidance and sex myoblast migration through extracellular matrix interactions .
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Structure: Contains inactive metalloprotease, disintegrin, and cysteine-rich domains critical for cell adhesion .
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Expression: Acts non-autonomously, suggesting indirect signaling roles in neuronal and migratory pathways .
No antibodies targeting UNC-71 are described in the literature. Research focuses on genetic interactions (e.g., with unc-6/netrin and integrins) rather than immunological tools .
EV71 Antibodies in Virology
Confusion may arise from the similarity between "UNC-71" and EV71 (Enterovirus 71), a pathogen linked to severe hand, foot, and mouth disease. Notable EV71-neutralizing antibodies include:
Broadly Neutralizing Antibody D5
Monoclonal Antibody 10D3
| Property | Details |
|---|---|
| Epitope | VP3 "knob" region (residues 59, 62, 67) |
| Specificity | Universal across EV71 subgenogroups (A, B2, B4, C2, C4) |
| Application | Diagnostic differentiation from coxsackievirus A16 (CVA16) |
UNC-Related Antibodies in Other Systems
While no UNC-71-targeting antibodies exist, UNC family proteins (e.g., netrin receptors UNC-5/UNC-40/DCC) have been studied in cancer and neurobiology. Example antibodies include:
| Antibody | Target | Application | Source |
|---|---|---|---|
| Anti-DCC | DCC receptor | Tumor suppression studies | Commercial kits |
| Anti-UNC5B | UNC5B | Schwann cell proliferation assays | Research tools |
Technical Considerations for Antibody Development
Hypothetical development of an anti-UNC-71 antibody would require:
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Immunogen Design: Recombinant UNC-71 disintegrin/cysteine-rich domains.
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Validation: Functional assays in C. elegans axon guidance models.
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Cross-Reactivity: Screening against human ADAM homologs (e.g., ADAM10/17).
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- Research suggests that UNC-71 operates in a non-cell-autonomous manner, influencing motor axon guidance and sex myoblast migration. It likely collaborates with integrins and UNC-6/netrin to provide axon guidance cues at specific decision points for motoneurons. PMID: 12783787
Q&A
Based on the analysis of patent documents and peer-reviewed studies, here is a structured FAQ addressing key research considerations for EV71-neutralizing antibodies (note: "unc-71" appears to be a typographical error; all available data relate to enterovirus 71 (EV71)):
How are EV71-neutralizing antibodies identified and validated in experimental systems?
Methodological approach:
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Plasmablast sorting: Isolate IgG+ plasmablasts from peripheral blood mononuclear cells (PBMCs) of convalescent patients (day 7 post-infection) using flow cytometry (CD19+CD3−CD20−CD38+CD27+) .
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Neutralization assays: Test antibody clones against multiple EV71 genotypes (B5, C4) using microneutralization tests with RD cells. Validate potency via IC<sub>50</sub> values (e.g., <10 ng/mL for high-potency clones) .
What factors determine antibody neutralization breadth across EV71 genotypes?
Key determinants:
How to resolve contradictions in serological vs monoclonal antibody neutralization data?
Resolution strategy:
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Perform antibody depletion assays using EV71 virion-coupled magnetic beads to quantify the contribution of specific epitopes to serum neutralization .
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Compare clonal dominance patterns via B-cell receptor sequencing (e.g., donors with >50% VP2 puff-specific clones show reduced cross-genotype neutralization) .
How to engineer bispecific antibodies for EV71 and coxsackievirus A16 (CVA16) cross-protection?
Protocol:
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Parental antibody selection:
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EV71-specific: CT11F9 (targets VP1 GH loop)
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CVA16-specific: NA9D7 (binds VP2 EF loop)
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Construct design: Use scFv-IgG format with EV71 scFv fused to heavy chain and CVA16 scFv to light chain .
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Validation:
What structural features enable IgM antibodies to outperform IgG in EV71 neutralization?
Mechanistic insights:
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Pentameric avidity: IgM 10D3 binds VP3 knob region with 10<sup>3</sup>-fold higher functional affinity than IgG analogs .
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Epitope accessibility: IgM’s larger footprint enables simultaneous engagement of VP1-219 and VP2-145 across adjacent protomers (cryo-EM reconstruction at 3.8 Å) .
How to optimize humanized antibodies for enhanced EV71 neutralization?
Humanization workflow:
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CDR grafting: Transplant murine CDRs (e.g., MA28-7) onto human framework regions (FRs) using Kabat numbering .
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Critical FR residues: Retain murine residues at positions 4L, 36L, 46L, 69H, 93H to maintain CDR conformation .
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Affinity maturation: Perform site-saturation mutagenesis at heavy chain position 103H (VP1 contact residue) .
