EXPB1 Antibody

FAQs for EXPB1 Antibody Research

Advanced Research Questions

• How to resolve contradictions in EXPB1’s IgE reactivity across plant species?
  Conflict: Arabidopsis EXPB1 shows no IgE reactivity, while grass homologs (e.g., Zea m 1) are potent allergens .
  Methodology:

  • Perform site-directed mutagenesis to swap key residues between EXPB1 and allergenic homologs.

  • Use surface plasmon resonance (SPR) to quantify IgE binding affinity .

  • Critical factor: A 12-residue divergent loop in domain 1 alters antibody accessibility .

• What methodologies map conformational epitopes on EXPB1?

  • Computational design: Graft complementary peptides onto antibody CDR loops to target disordered regions .

  • Molecular dynamics (MD): Simulate EXPB1-antibody complexes to predict binding interfaces .

  • Experimental validation: Use hydrogen-deuterium exchange mass spectrometry (HDX-MS) to identify protected regions upon antibody binding .
    Example: Two-loop DesAbs with cooperative binding peptides improve affinity by 5× .

• What challenges arise in crystallizing EXPB1-antibody complexes?
  Challenges:

  • EXPB1’s flexible glycan-binding groove complicates complex stabilization .

  • Antibody CDR loops may induce conformational changes in EXPB1 .
    Solutions:

  • Use single-domain antibodies (nanobodies) for reduced flexibility .

  • Employ cryo-EM as an alternative to X-ray crystallography for dynamic complexes .

Data Table: EXPB1 vs. Allergenic Homologs

Methodological Recommendations

• Antibody engineering: Use Rosetta-based design for CDR loop optimization .

• Data analysis: Leverage NGS tools (e.g., Geneious Biologics) for high-throughput antibody sequence clustering .

• Conflict resolution: Combine SPR, mutagenesis, and MD simulations to reconcile IgE reactivity data .