Humanization & Sequence Optimization

AtaGenix provides antibody humanization services to reduce immunogenicity of non-human antibodies while preserving binding affinity and specificity. We design 18 humanized sequence variants, express and characterize the top candidates, and deliver 3 optimized humanized antibodies with confirmed affinity — guaranteeing at least one retains affinity comparable to the parental chimeric antibody.

Humanized Sequences Designed

Best Candidates Delivered

~10 wks

Total Timeline

Guaranteed

Affinity Retention (≥1 variant)

Antibodies from mouse, rabbit, rat, or camelid sources often trigger anti-drug antibody (ADA) responses in humans. Humanization replaces non-human framework regions with human germline sequences while retaining the CDRs that determine antigen-binding specificity. AtaGenix uses CDR grafting combined with 3D structural modeling, back mutation prediction, and PTM risk analysis to ensure minimal affinity loss.

Humanization Types

Type	Description	Human Content
Chimeric	Non-human VH/VL fused to human constant regions (CH/CL)	~70% human (constant region only)
CDR-Grafted (Humanized)	Non-human CDRs grafted into human framework + back mutations at critical positions	~90–95% human
Fully Human	Derived from human libraries (phage display) or transgenic animals — no humanization needed	100% human

18 Designs, 3 Delivered

We design 18 humanized sequence variants exploring different human germline frameworks and back mutation combinations. All 18 are synthesized and expressed; the top 3 performers (by affinity and expression) are purified and delivered with full characterization data.

Affinity Retention Guarantee

We guarantee that at least one of the 3 delivered humanized antibodies retains affinity comparable to the parental chimeric antibody. Affinity is validated by SPR/BLI kinetic measurement (K_D, k_on, k_off) to quantify any changes.

Therapeutic-Grade Expertise

Our antibody engineering team includes scientists who have successfully developed approved therapeutic antibody drugs. We offer a complete downstream path: humanization → stable cell line development (24 weeks add-on) → up to 150L bioreactor production.

AtaGenix's antibody humanization service takes your non-human antibody sequence through CDR grafting onto high-homology human germline frameworks, 3D structural modeling for conformation-critical residue identification, back mutation prediction, and PTM/developability risk screening. We design 18 humanized variants, synthesize and express all of them, measure affinity for each, and deliver the 3 best candidates with purified protein and kinetic data. Guarantee: at least one retains affinity comparable to the chimeric antibody. Total timeline: approximately 10 weeks. Supported source species: mouse, rabbit, rat, camelid. Supported formats: IgG, scFv, Fab, VHH, bispecifics.

Antibody Humanization Workflow

A 6-step pipeline from your parental antibody sequence to 3 purified, affinity-confirmed humanized candidates. Total timeline: ~10 weeks.

Sequence Analysis

~2 weeks

CDR identification
Human germline selection
Sequence alignment

Humanization Design

2–4 weeks

CDR grafting onto human FW
3D modeling & back mutations
PTM risk screening
18 variants designed

Gene Synthesis & Expression

~2 weeks

Codon optimization
Vector construction
Transient expression (CHO/HEK)

Purification

~2 weeks

Protein A/G purification
SDS-PAGE QC
Yield assessment

Affinity Measurement

~3 weeks

SPR/BLI kinetics
K_D, k_on, k_off for all variants
Comparison vs. chimeric

Data & Delivery

~1 week

Top 3 candidates selected
Sequences + purified protein
Full technical report

Service Scope

✓ CDR grafting onto high-homology human germline	✓ 3D homology modeling & structural analysis
✓ Back mutation prediction for affinity retention	✓ PTM risk & developability screening
✓ 18 humanized sequence variants designed	✓ Gene synthesis, codon optimization, expression
✓ SPR/BLI affinity validation (K_D, k_on, k_off)	✓ 3 best candidates purified & delivered
✓ Source: mouse, rabbit, rat, camelid antibodies	✓ Formats: IgG, scFv, Fab, VHH, bispecifics
✓ Guarantee: ≥1 variant affinity comparable to chimeric	✓ Optional: stable cell line (24 wks) + 150L production

Deliverables

Deliverable	Details
3 Humanized Antibodies	Purified protein of the 3 best-performing humanized variants (from 18 designed)
Humanized Sequences	All 18 humanized VH/VL sequences with germline alignment report
Back Mutation Report	List of back mutations with structural justification for each
Kinetic Data	SPR/BLI data (K_D, k_on, k_off) for all expressed variants vs. chimeric control
Technical Report	Design rationale, expression data, QC, affinity comparison, and recommendations

Need upstream services? If your antibody sequence is locked in a hybridoma cell line, start with our hybridoma sequencing service. After humanization, proceed to CHO Stable Cell Line construction for manufacturing-ready production. If you want fully human antibodies without humanization, consider Phage Display from our human naïve library (LiAb-SFMAX™).

Frequently Asked Questions

Why 18 humanized sequences instead of just a few?

Different human germline frameworks accept the same CDRs differently. Some frameworks preserve binding better but may have PTM risks; others are structurally cleaner but require more back mutations. By designing 18 variants across multiple germline families and back mutation strategies, we explore a broad design space and identify the optimal combination of humanization level, affinity, expression, and developability. The top 3 are delivered with full data so you can make an informed selection.

What is the affinity guarantee?

We guarantee that at least one of the 3 delivered humanized antibodies retains affinity comparable to the parental chimeric antibody, as measured by SPR/BLI kinetics. "Comparable" means the K_D of the humanized variant is within the same order of magnitude as the chimeric control. If none of the 18 variants meets this criterion, we work with you on a remediation strategy at no additional design cost.

What is the difference between humanization and using a human naïve library?

Humanization starts with an existing non-human antibody (e.g., from mouse hybridoma) and engineers it to be ~90–95% human by CDR grafting. This preserves the proven epitope and binding mode. A human naïve library (like our LiAb-SFMAX™ phage display library) discovers antibodies that are 100% human from the start but may bind different epitopes or have different kinetics. Humanization is the right choice when you already have a well-characterized lead with a validated epitope; phage display is better when starting from scratch.

Can I go directly to stable cell line after humanization?

Yes. AtaGenix offers stable cell line development as a direct add-on service (~24 weeks) using our proprietary high-expression vectors with commercial development license. This creates a seamless path from humanized antibody sequence to manufacturing-ready cell line, including up to 150L bioreactor capability for clinical-grade material production.

Humanization success depends on the structural compatibility between the parental CDRs and the selected human germline framework. Some antibodies with unusual CDR conformations may require additional engineering. Affinity guarantee applies to SPR/BLI-measured K_D comparison with chimeric control. Quote-based pricing.