FTRC Antibody
Description
Overview of FTC-Conjugated Antibodies
FTC (fluorescein isothiocyanate) is a fluorescent dye commonly conjugated to antibodies for immunofluorescence microscopy, flow cytometry, and antigen detection. These antibodies retain their antigen-binding specificity while enabling visualization or quantification of target molecules.
Key Applications
Structure and Mechanism of FTC-Conjugated Antibodies
FTC conjugation typically occurs at lysine residues or Fc regions of antibodies without compromising their binding affinity. This modification enables:
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Direct fluorescence detection of antibody-antigen complexes.
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Multiplex analysis in flow cytometry when paired with other fluorophores.
Example:
The anti-ASL1/FTC antibody binds to the TL antigen on thymocytes, causing antigenic modulation (patching and capping) observed via fluorescence microscopy .
Antigenic Modulation Kinetics
In studies using anti-ASL1/FTC antibodies, modulation kinetics vary between cell types:
| Cell Type | Modulation Time (50% Complete) | Capping Efficiency |
|---|---|---|
| RADA1 cells | 20 minutes | 90% (after 4 hours) |
| Thymocytes | 55 minutes | 70% (after 4 hours) |
| Source: |
Flow Cytometry Applications
The TRA-1-85/CD147 Fluorescein-conjugated antibody (FAB3195F) is used to detect the TRA-1-85 antigen, expressed on almost all human cell types :
| Parameter | Specification |
|---|---|
| Clone | TRA-1-85 |
| Specificity | Human TRA-1-85/CD147 |
| Applications | Flow cytometry, immunofluorescence |
| Reactivity | Human PBMCs |
Fc Receptor-Related Antibody Engineering
While not directly related to FTC, Fc region modifications are critical for enhancing antibody effector functions (e.g., ADCC, complement activation). Key strategies include:
Half-Life Extension
Fc Glycoengineering
| Modification | Effect |
|---|---|
| Afucoylation | 50-fold increased ADCC via FcγRIIIa binding |
| Bisecting GlcNAc | Enhanced FcγRIIIa interaction |
Challenges and Considerations
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Interference in Assays: FTC-conjugated antibodies may interact with other fluorophores or chemical compounds. For example, tenofovir/emtricitabine (PrEP) does not interfere with VRC01 detection in ELISA assays .
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FcR Polymorphisms: Variability in FcγRIIIa (Val158Phe) impacts ADCC efficacy, necessitating population-specific antibody optimization .
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Off-Target Effects: Fc-engineered antibodies require rigorous testing to avoid unintended immune activation (e.g., cytokine release) .
Product Specs
Target Background
- Kinetic studies indicate that while FTR (ferredoxin-thioredoxin reductase) reduces all ten chloroplast thioredoxin (Trx) isoforms, the efficiency of electron transfer varies significantly. Trx-m4 reduction occurs at a higher rate compared to other Trx-m isoforms. PMID: 28246333
- FTR is essential for plastid-encoded RNA polymerase (PEP)-dependent plastid gene expression during early chloroplast development. Specifically, FTRc is crucial for leaf greening, particularly in young tissues. PMID: 24890758
Q&A
Here’s a structured FAQ collection for academic researchers focused on FTRC Antibody (assuming this refers to IGF1R-targeting antibodies as described in the search results):
Advanced Research Challenges
Q4: How can conflicting data on IRS-1 phosphorylation as a predictive biomarker be resolved?
Analytical framework:
Q5: What computational approaches enhance antibody design against flexible binding regions?
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Use RFdiffusion models (fine-tuned for antibody loops) to generate single-chain variable fragments (scFvs) with improved flexibility .
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Validate AI-designed antibodies via surface plasmon resonance (SPR) and cryo-EM for structural accuracy .
Q6: How do combination therapies impact IGF1R antibody efficacy in resistant tumors?
Synergy strategies:
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Co-administer TLR8 agonists or PD-1 inhibitors to counteract immunosuppressive microenvironments .
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Test sequential dosing (e.g., IGF1R antibody followed by chemotherapy) to exploit metabolic vulnerabilities.
Methodological Pitfalls & Solutions
Q7: Why might in vitro binding data fail to predict in vivo efficacy?
Common limitations:
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Lack of stromal interactions in monolayer cultures.
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Solution: Use 3D organoid models or patient-derived xenografts (PDXs) with intact tumor microenvironments.
Q8: How should researchers address batch-to-batch variability in antibody production?
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Implement mass spectrometry-based peptide mapping for lot consistency.
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Establish functional potency assays (e.g., neutralizing activity in IGF1-dependent cell lines).
Emerging Techniques & Validation
Q9: What orthogonal methods confirm target engagement in vivo?
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Perform immunoPET imaging with zirconium-89-labeled antibodies.
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Analyze phospho-IGF1R/IRS-1 levels in tumor lysates via targeted mass spectrometry.
Q10: How can AI-driven antibody design accelerate discovery pipelines?
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RFdiffusion-generated scFvs achieve moderate affinity (K<sub>D</sub> ~10<sup>-7</sup> M) without experimental optimization .
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Refinement via yeast display or phage libraries can improve affinity to <10<sup>-9</sup> M.
