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Antibody Humanization Antibody Humanization

Antibody Humanization

Antibody Optimization
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Humanized antibodies with guaranteed affinity preservation in just 3-4 weeks.

Our antibody humanization service addresses the challenge of immunogenicity by employing CDR grafting onto selected human germline frameworks. Combined with limited back mutations to maintain loop conformation and antigen affinity, we deliver antibodies with reduced anti-drug antibody (ADA) risk and reliable performance. Our integrated bioinformatics and structural modeling platform ensures precise germline selection and sequence optimization, while minimizing the number of framework modifications. With a guaranteed affinity drop of no more than three-fold compared to the parental antibody, and a typical turnaround of 3–4 weeks, we provide a fast and dependable path for optimizing antibody candidates.

Our antibody discovery services also include:

Antibody Humanization Overview

Integrated Workflow from Sequence Design to Affinity Testing

Our workflow incorporates advanced antibody structure simulation and optimization tools to guide germline selection and CDR grafting. In addition to sequence design, we include risk assessment of post-translational modifications (PTMs) to minimize liabilities during development. Each project progresses through codon optimization, gene synthesis, transient expression, purification, and stringent quality control, followed by binding validation using Biacore SPR analysis. This integrated strategy ensures that humanized antibodies not only maintain comparable affinity to their parental versions but also demonstrate favorable biochemical properties for downstream development.

Antibody Humanization Overview

Advantages of our Antibody Humanization Service

Guaranteed Affinity

  • We offer guaranteed affinity (drop less than 3 folds) compared to the parental antibody
  • Less than 5 back mutations for VH/VL each and 7 for VHH

Quick Turnaround Time

  • We deliver humanized antibodies in just 3-4 weeks
  • 10+ years of experience and expertise in humanized antibody design

Optimal Functionality

  • Receive an affinity detection report with Biacore
  • We can validate your humanized antibodies with FACS (fluorescence-activated cell sorting) flow cytometry

Demonstrated Success with Minimal Back Mutations

Biointron’s platform has been applied successfully to antibodies from multiple species and formats, including IgG and Fab fragments from mice and rabbits, as well as VHH domains from camelids and sharks. Our approach limits back mutations to fewer than five for VH/VL and fewer than seven for VHH, providing efficient grafting with minimal framework modification. This controlled approach is important because excessive back mutations can compromise the purpose of humanization by reintroducing non-human residues, potentially increasing immunogenicity or destabilizing the antibody scaffold. By carefully selecting only the residues essential for maintaining CDR conformation and antigen binding, we achieve a balance between reducing immunogenic risk and preserving functional affinity.

Antibody Humanization Workflow

Gene Synthesis

Plasmid Construction and Preparation

Transient Expression

Affinity Purification

Stringent Quality Control

Affinity Ranking

Antibody Humanization Service Timeline

Service Step Service Description Timeline Deliverables
Design of humanized antibody
  • Bioinformatics analysis
  • 3D structure modeling and identification of back mutations
  • Human germlines selection
  • In silico CDR-grafting
  • Sequence optimization
2-3 days
  • Designed humanized antibody sequences
Humanized antibody recombinant production variants 9 to 25
  • Codon optimization and gene synthesis
  • Subcloning into expression vector
  • Transient expression
  • Purification
  • QC analysis
2 weeks
  • Purified humanized antibody samples for in house test
Affinity Ranking
  • Affinity Determination with soluble antigen via Biacore 8K
1 week
  • Detailed report
  • Design of humanized antibody
    Timeline

    2-3 days

    Description
    • Bioinformatics analysis
    • 3D structure modeling and identification of back mutations
    • Human germlines selection
    • In silico CDR-grafting
    • Sequence optimization
    Deliverables
    • Designed humanized antibody sequences
  • Humanized antibody recombinant production variants 9 to 25
    Timeline

    2 weeks

    Description
    • Codon optimization and gene synthesis
    • Subcloning into expression vector
    • Transient expression
    • Purification
    • QC analysis
    Deliverables
    • Purified humanized antibody samples for in house test
  • Affinity Ranking
    Timeline

    1 week

    Description
    • Affinity Determination with soluble antigen via Biacore 8K
    Deliverables
    • Detailed report

Case Study

Case 1:

The affinity of humanized antibody (measured by SPR, Biacore 8K) is comparable with the parental antibody.

Antibody A
Antibody A
Humanized Antibody A
Humanized Antibody A
The affinity of humanized antibody is comparable with the parental antibody.

Case 2:

The parental antibody is VHH. Based on the SPR results, the affinity of humanized antibodies is similar to that of the parental antibody.

Antibody Humanization Case 2
"As a scientist at Biointron, my team’s collaborative spirit and shared commitment to excellence creates a positive and dynamic environment where I am encouraged to work to the best of my ability. Combined with cutting-edge technologies, you can be confident in the quality of our antibody humanization service.”
Jiwen Du
Jiwen Du
Antibody Optimization Team

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