In Silico Design
3D Structural Modeling
5+ Prediction Algorithms
≥85% Humanization
1–2 Week Turnaround
Service Overview
Therapeutic antibodies derived from non-human species — most commonly rodents — carry immunogenic framework regions that can trigger anti-drug antibody (ADA) responses in patients, limiting efficacy and tolerability. Humanization replaces non-human framework residues with their human germline counterparts while preserving the antigen-binding complementarity-determining regions (CDRs), yielding a molecule that is immunologically indistinguishable from a fully human antibody at the framework level. Traditional CDR grafting approaches often sacrifice binding affinity or introduce manufacturability liabilities in the process. ICDMO's in silico humanization platform addresses this challenge by integrating three-dimensional structural analysis, multi-parameter immunogenicity prediction, and physics-based free-energy calculations to design humanized antibodies that simultaneously optimize humanness, binding potency, biophysical stability, and production fitness — before a single experiment is run. Our computational workflow is grounded in rigorous structural biology and supported by a validated set of bioinformatics tools for epitope mapping, surface characterization, and post-translational modification (PTM) risk assessment. Each design deliverable includes a comprehensive analysis report and a ranked set of humanized sequence variants ready for gene synthesis and expression testing.
Key Features & Advantages
Humanization Percentage
Framework humanness maximized to ≥85% (Kabat/IMGT) while preserving CDR loop conformation and canonical geometry.
CDR Immunogenicity
T cell, B cell, and MHC II epitope burden within CDR loops minimized using five validated prediction algorithms in parallel.
Antigen Binding Affinity
Binding affinity retention constrained to ≥50% vs. parental antibody by MM-GBSA free-energy estimation on the Fv–antigen complex.
Expression & Manufacturability
Manufacturability flags (aggregation-prone regions, unpaired cysteines, rare codons) resolved to support robust CHO/HEK expression.
Aggregation Potential
Hydrophobic and aggregation-prone surface patches identified and eliminated or minimized to improve formulation stability.
Thermodynamic Stability
Thermodynamic stability (Tm) and long-term colloidal stability maintained or improved relative to the parental antibody.
Sequence Integrity (PTM)
Deamidation, oxidation, glycosylation and proteolysis liability sites mapped and de-risked throughout framework and CDRs.
Bioinformatics Analysis Suite
1PTM analysis — deamidation, oxidation, glycosylation, and proteolysis motif mapping onto 3D model
2Aggregation surface analysis — identification and scoring of hydrophobic surface clusters
3Electrostatic patch analysis — positive, negative, and hydrophobic patch characterization
4T cell epitope prediction using AMPHI and Rothbard–Taylor algorithms
5B cell epitope prediction using DNASTAR algorithm
6MHC II epitope prediction using Sette algorithm
7Antigenicity scoring using Jameson–Wolf and Welling methods
8Humanization percentage calculation (Kabat / IMGT definitions)
Back-Mutation Decision Framework
Structural Integrity Criterion
▸Substitution disrupts an ionic bond, hydrogen bond, or hydrophobic interaction with a CDR residue
▸Substitution affects a canonical residue, VH/VL interface residue, or Vernier zone residue
▸Substitution introduces steric clash or alters CDR canonical loop geometry
Biophysical Liability Criterion
▸Substitution measurably increases predicted immunogenicity epitope burden
▸Substitution increases aggregation-prone surface area or positive patch intensity
▸Substitution introduces a new 3D PTM motif (deamidation, oxidation, glycosylation site)
▸Substitution causes significant shift in 3D isoelectric point (pI)
ICDMO Quality Standards
Standard 1 — Humanization ≥85%
▸The proportion of human germline residues in combined VH/VL framework regions must reach ≥85% by Kabat/IMGT definition
▸This level of humanness is associated with substantially reduced anti-drug antibody (ADA) risk in clinical use
▸All delivered variants are scored and reported against this threshold before recommendation
Standard 2 — Binding Retention ≥50% vs. Parental
▸Predicted binding free energy of the humanized design must not exceed a 2-fold reduction vs. parental antibody
▸Computationally assessed by MM-GBSA free-energy estimation on the ternary Fv–antigen complex
▸Subsequently confirmed by wet-lab SPR or BLI experiments on expressed humanized variants
Deliverables
1Ranked humanized VH/VL sequence variants in FASTA format
2Comprehensive 3D structural analysis report with annotated Fv models
3Immunogenicity epitope comparison — parental vs. humanized (all algorithms)
4Surface patch and aggregation risk assessment with visual maps
5PTM liability mapping for each proposed variant
6Detailed back-mutation rationale document with structural justification
Service Workflow
1
3D Antibody Modeling
Build a high-accuracy homology model of the parental antibody Fv/Fab domain from VH/VL sequence input. Identify CDR loops, canonical residue positions, and CDR–antigen contact residues that must be protected during CDR grafting.
2
Immunogenicity Epitope Analysis
Predict T cell, B cell, and MHC II immunogenic epitopes on both the parental sequence and the 3D structural surface using five validated bioinformatics algorithms (AMPHI, Rothbard–Taylor, DNASTAR, Sette, Jameson–Wolf).
3
Surface & PTM Liability Analysis
Characterize positive, negative, and hydrophobic electrostatic patches. Score aggregation-prone surface regions. Map all PTM liability sites (deamidation, oxidation, glycosylation, proteolysis) with side-chain accessibility scoring.
4
Human Germline Acceptor Selection
BLAST search IMGT and NCBI germline databases to identify VH and VL germline genes with highest sequence identity to the parental framework. Assess 3D structural compatibility to ensure selected acceptor accommodates parental CDR conformations.
5
CDR Grafting & Back-Mutation Design
Graft parental CDRs onto the selected human germline acceptor framework. Apply the back-mutation decision criteria to each non-human → human position. Deliver a ranked panel of optimized humanized variants satisfying both quality standards.
Service Process
1
Online Consultation
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2
Solution Matching
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3
Service Contract
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4
Project Execution
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5
Data Delivery
Specifications & Turnaround
Note: All services are for research use only and not for diagnostic or therapeutic purposes. For GMP-grade services, please contact us separately.
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Contact Us Online ConsultationStandard Deliverables
✓Sequence-verified final product
✓Full QC documentation
✓Certificate of Analysis (CoA)
✓Electronic data files
✓Project completion report
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