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Antibody Humanization

Our Approach

Leveraging large-scale human framework libraries and AI structure prediction, matching optimal framework regions from a wider sequence space, precisely maintaining CDR-framework interactions — no back-mutations needed, higher humanization, shorter timelines.

Validated Results

90-95%
Humanization Level
0
Back-mutations
20 / 100
Per Round
4h
Computation Time

Why Is This Needed?

"Murine monoclonal antibodies face significant challenges in clinical applications from immunogenicity (HAMA reactions) and insufficient effector functions. Humanization eliminates heterologous features and enhances ADCC/CDC effector functions — a core strategy for improving therapeutic efficacy and safety."

Key Advantages

Key Advantages

Traditional Limitations

Limited Framework Selection

Traditional CDR grafting is limited to highly homologous frameworks, unable to screen from a larger space for the truly optimal acceptor framework.

Affinity Easily Lost

Grafting often causes affinity decline due to CDR-framework incompatibility, requiring multiple rounds of back-mutation trial and error.

Long Cycles, High Costs

Extensive expression, purification, and screening are time-consuming, relying on experience-based mutation design.

Our Advantages

Intelligent Diversified Framework Matching

Breaking through traditional similar-framework limitations, exploring greater sequence space via massive human framework libraries, providing diversified framework candidates for subsequent molecular optimization.

Structure-Driven Affinity Retention

AI-predicted antibody structures accurately model CDR-framework interactions, which is key to maintaining affinity after humanization.

No Back-Mutations, High Success Rate

No back-mutations needed — typically only one round of expressing 10~20 antibodies yields candidates with affinity and expression comparable to parental, significantly reducing time and experimental costs.

Multi-Dimensional Candidate Screening

Scores, clusters, and screens candidates based on humanization level, stability, and predicted biophysical properties, outputting optimized recommendations.

Validation

Case Studies

In a collaboration with an international pharmaceutical client, AI humanization was performed on 2 murine monoclonal antibodies Ab2 and Ab3. Based on AI prediction results and model scoring, 10 humanized antibodies were selected for recombinant expression to measure expression levels and affinity.

Designing just 10 candidates yielded at least 1 antibody outperforming the patent-reported molecule, with 90%-95% humanization

All antibodies showed expression levels comparable to or higher than parental; in one system, 3 antibodies had higher affinity than parental, others within 3-fold difference, with 1 antibody exceeding the patent-reported humanized antibody in affinity; humanization level reached 90%-95%.

Parental
Patent
Click.mAb.
Sequence Identity (%)707580859095100Heavy ChainLight Chain
Humanization Level (Ab2)
Parental
Patent
Click.mAb.
Sequence Identity (%)707580859095100Heavy ChainLight Chain
Humanization Level (Ab3)
Parental
Patent
Click.mAb.
KD (M)1e-91e-81e-71e-61e-5Ab2Ab3
Affinity KD (M)
Parental
Patent
Click.mAb.
Expression Level (mg)00.20.40.60.81Ab2Ab3
Expression Level (4ml scale)

Pipeline

Computational Pipeline

Fully automated AI humanization workflow, from sequence submission to recommended results in one step.

Input & Preprocessing
Structure Modeling & Ab Analysis
Humanization & Framework Design
Library Generation, Scoring & Screening
Recommendation

Submit VH/VL sequences → Structure modeling & CDR identification → AI framework design → Generate 10,000–100,000 variants with scoring & screening → Output recommended sequences, Germline alignment reports & risk site scoring

Report

Report Examples

Below are partial screenshots from the antibody humanization service delivery report, showing the complete workflow from sequence analysis to humanization scheme evaluation.

Use Cases

Use Cases and Deliverables

Best suited for humanization optimization before downstream development of murine mAbs.

4h2,888 credits / antibody / run

Best-Fit Scenarios

Murine mAbs need optimization before development advancement
You need to reduce immunogenicity risk while preserving affinity and functional activity as much as possible
You need a ranked set of humanized candidates for expression validation and backup screening

Inputs to Prepare

Antibody VH/VL sequences

What You Receive

Antibody humanization report
Top 20 & 100 candidate sequences ranked by clustering score
Germline type analysis
Risk-site prediction and detailed information

What is the advantage over traditional CDR grafting?

Traditional CDR grafting often leads to decreased affinity, frequently requiring multiple rounds of back-mutation. The platform leverages a large-scale human germline framework library and structural compatibility assessment to identify framework combinations better suited for the parent CDRs, eliminating the need for back-mutation.

How does the platform preserve affinity while improving humanness?

Affinity retention depends on the 3D support relationship between CDRs and frameworks. The platform evaluates Vernier regions, CDR interfaces, and key framework sites to avoid disrupting CDR conformation after framework replacement.

Why is “no back-mutation” valuable?

Back-mutations usually mean multiple empirical trial-and-error rounds and extra experimental cost. If structure-driven framework matching directly yields candidates with affinity and expression close to the parental antibody, validation cycles can be significantly shortened.

Why is multi-dimensional screening better than only checking humanness?

Humanness percentage alone can miss stability, risk sites, and expression concerns. The platform combines humanness, structural stability, predicted biophysical properties, and clustering scores to output top candidates for downstream experimental selection.

Start Your AI-Driven Humanization Journey

Overcome immunogenicity barriers while maintaining affinity and achieving significant improvements in humanization levels.