Antibody Humanization Antibody Humanization

Antibody Humanization

Antibody Optimization
  • Guaranteed Affinity Comparable to Parental Antibody
  • Quick Turnaround Time of 3-4 Weeks
  • Suitable for IgG, scFv, Fab, VHH, and more
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Antibody Humanization Overview

Remarkable advancements have taken place in recombinant antibody sciences, and made possible significant clinical success of therapeutic antibody drugs. However, major challenges remain, such as the anti-drug antibody (ADA) issue associated with the immunogenicity of non-human antibodies. This response occurs when human immune systems react to the foreign protein, leading to moderate to serious safety and efficacy concerns. To tackle this issue, chimeric antibodies were created, where the antibody’s variable domains were recombined with human constant domains, thus preserving antibody specificity.1 However, the problem of immunogenicity-triggered liabilities needs to be further targeted.

Antibody Humanization Overview

Antibody humanization is a tool that was developed to better tackle the limitation posed by ADA by genetically modifying the non-human antibodies to be more of human source. Principally, the strategy involves CDR (complementarity determining region) grafting (non-human antibodies are grafted to human frameworks of high homology) and back mutations (some parental residues in framework regions are retained as they have been shown to affect CDR loop conformation and antibody affinity2). The resulting humanized antibodies carry lower immunogenicity risk than parental or chimeric antibodies.3


Exploiting Biointron’s antibody structure simulation and optimization platform, we have been able to identify the best human germline sequence templates, and to accomplish CDR crafting and back mutations in such a way that maximally keep antibodies’ affinity properties intact, we also include risky post-translational modifications (PTMs) analysis into our humanization strategy to fulfill extra antibody optimization.

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
  • If the specified affinity is not attained, there will be no cost

Quick Turnaround Time

  • We deliver humanized antibodies in just 3-4 weeks
  • 10+ years of experience and expertise in humanized antibody design
  • Successful production of more than 20,000 antibodies

Optimal Functionality

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

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

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

Antibody Humanization Service FAQs

References

  • Pornnoppadol, G., Zhang, B., Desai, A. A., Berardi, A., Remmer, H. A., Tessier, P. M., & Greineder, C. F. (2021). A hybridoma-derived monoclonal antibody with high homology to the aberrant myeloma light chain. PLOS ONE, 16(10), e0252558. https://doi.org/10.1371/journal.pone.0252558
  • Safdari, Y., Farajnia, S. (2013). Antibody humanization methods – a review and update. Biotechnology and Genetic Engineering Reviews, 29(2), pp. 175–186. https://doi.org/10.1080/02648725.2013.801235
  • Harding, F. A., Stickler, M. M., Razo, J., & DuBridge, R. B. (2010). The immunogenicity of humanized and fully human antibodies: Residual immunogenicity resides in the CDR regions. MAbs, 2(3), 256-265. https://doi.org/10.4161/mabs.2.3.11641
  • Lu, RM., Hwang, YC., Liu, IJ. et al. Development of therapeutic antibodies for the treatment of diseases. J Biomed Sci 27, 1 (2020). https://doi.org/10.1186/s12929-019-0592-z

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