scpl-4 Antibody

What experimental methods validate CXCL4 antibody reactivity in autoimmune diseases?

Methodological approach:

• Use homemade ELISA to quantify anti-CXCL4 IgG/IgA in serum ( ):

  • Compare against healthy donors (HD) and disease controls (e.g., SLE, ulcerative colitis)

  • Validate with Western blot using recombinant CXCL4

• Clinical correlation: Measure IFN-α levels via ELISA and interferon-responsive genes (e.g., MX1) in blood/skin biopsies

Key data:

How do CXCL4-DNA complexes influence interferon signatures?

Experimental design:

• Isolate circulating immune complexes (CICs) from SSc patient plasma

• Use immunofluorescence to colocalize CXCL4 and DNA in skin biopsies

• Stimulate pDCs with CXCL4-DNA complexes and quantify IFN-α via Luminex

Critical finding: CXCL4-DNA complexes correlate with:

• Blood IFN-α levels (r = 0.72, p < 0.001)

• Skin MX1 expression (r = 0.65, p = 0.004)

How to assess T-cell help in anti-CXCL4 antibody production?

Protocol:

• Isolate PBMCs from SSc patients and perform BrdU proliferation assays:

  • Stimulate CD4+ T-cells with 10 μg/mL recombinant CXCL4 for 5 days

  • Gate on CD3+CD4+BrdU+ populations (Figure S2 in )

• Statistical correlation: Anti-CXCL4 antibody titers vs. T-cell proliferation index (r = 0.81, p = 0.003)

Implication: 38% of SSc patients show CXCL4-specific T-cell responses, suggesting linked recognition between B and T cells .

What computational methods predict antibody developability?

Framework:

• Train GAN models on 31,416 human antibody sequences with optimal biophysical properties

• Generate 100,000 in silico variable regions filtered by:

  • Humanness score >90%

  • Hydrophobicity index <0.35

  • Thermal stability >68°C (DSC)

• Validate top candidates via:

  • CE-SDS for purity

  • HIC-HPLC for aggregation propensity

Validation results (n=51):

How to resolve conflicting data on CTLA-4 blockade in CXCL4-related models?

Contradiction analysis:

• Issue: Anti-CTLA-4 antibodies may target both effector T-cells and T-regs ( )

• Experimental reconciliation:

  • Use humanized CTLA-4 mice to isolate compartment-specific effects

  • Compare tumor growth in B16/BL6 melanoma models with:

    • Unicompartmental (T-reg only) vs. bicompartmental blockade

  • Key metric: Tumor volume reduction = 68% with bicompartmental vs. 22% unicompartmental (p < 0.001)

Methodological Recommendations

• For B-cell epitope mapping: Combine hydrogen-deuterium exchange mass spectrometry (HDX-MS) with alanine scanning mutagenesis of CXCL4

• In cytokine storm models: Monitor CXCL4-CCR1 axis using phosphoflow cytometry (CD4+pSTAT5)

• Multi-omics integration: Pair single-cell RNA-seq of CXCL4+ macrophages with spatial transcriptomics in fibrotic tissues