GOS11 Antibody

Basic Research Questions

How to establish target specificity for newly developed antibodies?

• Method: Combine orthogonal validation techniques:

  • Western blot comparison between human/mouse orthologs (e.g., 80% identity threshold for species-specificity)

  • Competitive ELISA with recombinant protein variants

  • Knockout validation using CRISPR-modified cell lines

What experimental controls are essential for neutralizing antibody assays?

• Required controls:

  Control Type	Purpose	Example from Literature
  Isotype control	Baseline signal correction	Mouse IgG1 in SFTSV neutralization
  Antigen-negative cells	Specificity verification	Uninfected Vero cells in IFA
  Knockout validation	Target confirmation	IL-11−/− mice in IHC staining

Advanced Research Questions

How to resolve discrepancies between in vitro neutralization and in vivo efficacy?

• Troubleshooting workflow:

  • Confirm target engagement via SPR/BLI (≥1 nM affinity recommended)

  • Assess pharmacokinetics using ultra-sensitive SP-X assays (LLOQ: 0.006 pg/mL)

  • Evaluate Fc-mediated effector functions (ADCC/CDC activity)

What computational strategies improve antibody epitope precision?

Key Technical Challenges from Literature

Data Contradiction Analysis Protocol

When facing conflicting results (e.g., strong in vitro neutralization vs. poor in vivo protection ):

• Verify antibody integrity:

  • SEC-HPLC for aggregation (<5% aggregates required)

  • SPR confirmation of on-rate (kon > 1×10^5 M−1s−1)

• Model species limitations:

  • Compare FcγR polymorphisms between human/A129 mice

  • Use humanized Fc variants in murine models