ResearchAlphaFold2pLDDTValidationStructure Prediction

Structure Validation with AlphaFold2: Metrics, Thresholds, and Ranking

Every candidate design generated by Foldexa is validated using AlphaFold2 structure prediction. This article explains our scoring metrics — pLDDT, iptm, PAE — the Foldexa Score formula, and how we rank candidate outputs from both pipelines.

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Azamat Armanuly

CEO & Bioengineer, KAIST

Feb 28, 2026 7 min read

Every design generated by Foldexa — whether from DiffAb or RFdiffusion — must pass structural validation before being shown to you. Here is exactly how that validation works.

AlphaFold2 Output Metrics

AlphaFold2 (and AlphaFold2 Multimer for complexes) produces four key numerical outputs beyond the predicted 3D structure itself. Foldexa uses three of them in its validation pipeline.

MetricRangeFoldexa UseNotes
pLDDT0–100Primary filterPer-residue confidence; > 85 is high confidence
iptm0–1Complex confidenceInterface predicted TM-score; > 0.7 is good
PAE0–35 ÅInterface qualityPredicted Aligned Error; lower = more confident
ptm0–1Internal checkGlobal chain confidence; not exposed in UI

pLDDT: Per-Residue Confidence

pLDDT (predicted Local Distance Difference Test) measures how confident AlphaFold2 is in each residue's position relative to its neighbors within 15 Å. It is not a measure of how well the structure will match an experimental structure — it is a measure of internal consistency. A protein that is predicted to be intrinsically disordered will have low pLDDT even if the prediction is correct.

Common Misconception

High pLDDT does not mean the protein folds in reality — it means AlphaFold2 is internally consistent about the predicted structure. A well-packed, globular design will naturally score higher than a flexible loop region. Always consider the context: CDR loops legitimately score lower than framework regions.

pLDDT RangeTierFoldexa Interpretation
≥ 90Very HighBackbone highly reliable; suitable for structure-guided downstream design
70 – 89HighGood confidence; acceptable for most applications
50 – 69LowPossibly disordered or low-confidence region; review manually
< 50Very LowPotentially unstructured; design likely does not fold as intended

The Foldexa Score Formula

Individual metrics are combined into a single Foldexa Score for ranking. We use a weighted composite that emphasizes interface confidence for complex designs:

Foldexa Score formula
def foldexa_score(plddt_mean: float, iptm: float, pae_interface_mean: float) -> float:
    """
    Compute Foldexa Score (0–1).

    Args:
        plddt_mean: Mean pLDDT of CDR or scaffold residues (0–100)
        iptm: AlphaFold2 Multimer iptm score (0–1)
        pae_interface_mean: Mean PAE across interface residue pairs (Å)

    Returns:
        Foldexa Score between 0 and 1
    """
    plddt_norm = plddt_mean / 100.0
    pae_norm = max(0.0, 1.0 - pae_interface_mean / 50.0)

    return 0.40 * plddt_norm + 0.40 * iptm + 0.20 * pae_norm

Tier System

TierFoldexa ScoreApprox. PercentileRecommendation
S≥ 0.85Top 5%Prioritize for wet-lab synthesis
A0.75 – 0.84Top 20%Strong candidates; validate before synthesis
B0.65 – 0.74Top 40%Acceptable; use for backup or diversity
C< 0.65Bottom 60%Flagged; review manually or discard

Practical Guidance

For a typical 50-design campaign, expect 4–7 S-tier designs, 8–12 A-tier, and the rest in B or C. Ordering 3–5 S-tier designs for expression is a cost-effective starting point — enough diversity to hedge against expression failure.

Reading the Results JSON

Completed job result structure
{
  "job_id": "job_9f3a2b",
  "status": "completed",
  "designs": [
    {
      "design_id": "d001",
      "rank": 1,
      "tier": "S",
      "foldexa_score": 0.912,
      "plddt_mean": 92.4,
      "iptm": 0.91,
      "pae_interface_mean": 4.2,
      "sequence": "EVQLVESGGGLVQPGGSLRL...",
      "pdb_url": "https://results.foldexa.com/job_9f3a2b/d001.pdb",
      "passed_self_consistency": true,
      "self_consistency_rmsd": 0.74
    }
  ],
  "summary": {
    "total_designs": 50,
    "passed_plddt_85": 44,
    "tier_s": 5,
    "tier_a": 11,
    "tier_b": 18,
    "tier_c": 16
  }
}
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Written by

Azamat Armanuly

CEO & Bioengineer, KAIST