Home » Biologics Discovery » Antibody Lead Optimization » Antibody Humanization

Service Introduction

Express Service: As Fast as 2.5 weeks to Get Your Antibody Humanized

Through CDR grafting and back mutation technologies, the express package can be used to produce humanized antibodies with high expression levels, good thermal stability, and comparable affinity with the parental antibodies as fast as 2.5 weeks.

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Deluxe Service: Get Humanized Antibody with No Affinity Loss

Deluxe package can be used to obtain the humanized antibodies without affinity loss through CDR grafting, back mutation, and FASEBA high-throughput screening technology as fast as 8 weeks.

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FASEBA is a patented E.Coli based system that selects good affinity, expression level and thermostable candidates. It is used to select the best protein binders from a large number of protein candidates in a high throughput fashion, without actually purifying these proteins

Each saturation mutation library is transformed into FASEBA E.coli based expression system

Antibody Humanization

Immunogenicity can potentially lead to reduced efficacy through rapid clearance or neutralization of the drug, or toxicity due to cross-reaction with proteins in the body, which can have catastrophic consequences for the patient. Antibody humanization & antibodies characterization can reduce the immunogenicity of monoclonal antibodies derived from xenogeneic sources and to improve their activation of the human immune system by replacing non-human antibody frameworks with human ones. It is a very important step in therapeutic antibody discovery process.

A common method for humanization of non-human antibodies is complementary determining regions (CDR) grafting in which the CDRs of non-human antibodies are grafted onto the human frameworks. But only CDR grafting is not enough to get eliminate the risk of immunogenicity very well. Usually, back mutation will be performed to further transfer CDRs into human frameworks as similar as possible. In this step PTM hot spot analysis can be applied to increase the probability to get humanized antibody with least affinity loss and normal function.

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Also, FASEBA(Fast Screening for Expression, Biophysical-properties and Affinity) high throughput screening can be applied to screen out humanized antibody with best affinity.

ProBio provides 2 packages for antibody humanization service. Deluxe package using CDR-grafting, back mutation with PTM analysis and screening with FASEBA which can guarantee no affinity loss for the humanized antibody. Express package using CDR grafting and back mutation to generate humanized antibody as fast as 3 weeks.

Patented FASEBA screening platform

  • scFv, Fab and various formats humanized
  • 3-D structural modeling
  • Patented screening platform to screen affinity, thermostability, expression level & epitope, no royalty burden
  • Guaranteed delivery of best humanized abs with comparable affinity and good expression level & thermostability
  • Possible to combine with affinity maturation to further improve the affinity by 5-10 fold in <6 months
  • Antibody Humanization

    • 400+ ab humanization projects delivered
    • All abs showed at least comparable affinity with parental ab
    • >1/3 of humanized ab showed higher affinity than parental ab

How to Avoid Developability Risk in the Early Stage of Discovery?

We offer 3 developability assessment packages for antibody drug candidates at different discovery stages to identify the potential risks of manufacture and clinical failure.

Service Package

Deluxe Humanization --- Back mutaiton library screening with FASEBA

  Milestone Deliverables Turnaround
1 Chimeric antibody production and Binding confirmation
  • Sequence, DNA and chimeric ab & top 3 humanized abs (0.5 mg)
  • Guarantee no affinity loss
11 weeks
2
  • Developability Prediction:PTM analysis,Immunogenicity prediction,Aggregation analysis
  • Back mutation library design and library construction
3 Humanized antibody selection
4 Humanized antibody production and characterization
5 Affinity maturation(Optional)
  • Guaranteed 5-10 fold affinity higher than parental ab
Additional
8-9 weeks

The turnaround time listed here is the time required for the service itself. For international shipping, please refer to the time provided by PM during project discussion.

Express plus humanization --- CDR grafting with back mutation

  Milestone Deliverables Turnaround
1 Chimeric antibody production and binding confirmation
  • Sequence, DNA and chimeric ab & top 3 humanized abs (0.5 mg)
  • Guarantee at least 1 ab with affinity comparable with parental ab
12 workdays
2 Developability Prediction(optional):PTM analysis Immunogenicity prediction Aggregation analysis
3 Express humanization by rational design and sequence synthesis
4 Humanized antibody production and affinity ranking
5 Humanized antibody production and affinity determination

The turnaround time listed here is the time required for the service itself. For international shipping, please refer to the time provided by PM during project discussion.

  • Human framework diversity
  • Fast turnaround time
  • Guaranteed delivery of humanized abs with comparable affinity with parental abs

Case Study

  • samples Chi² (RU²) ka (1/Ms) kd(1/s) KD (M) Rmax (RU)
    humanized antibody 5.03E-02 3.67E+05 6.90E-05 1.88E-10 70.5
    parental antibody 2.11E-01 4.02E+05 5.56E-05 1.38E-10 73.6

    Table.1 Affinity kinetic analysis

  • Fig.1 Affinity kinetic analysis of humanized antibody

  • ELISA test was performed to evaluate binding between humanized and chimeric antibodies, and the same binding profile was observed based on the ELISA results.

  • Fig.2 Comparison of the affinity of humanized antibodies to parental antibody

  • FACS test was performed to evaluate binding between humanized and chimeric antibodies, and the same binding profile was observed based on the FACS results.

  • Fig.3 Comparison of the affinity of humanized antibodies to parental antibody

Service features

  • Extensive experience:
        20-year experience in antibody engineering
        Successfully performed 400+ antibody humanization projects
  • Package Feature:
        Express Plus Package: 3 weeks to deliver desired humanized antibody
        Deluxe Package: less immunogenicity issues
  • Guaranteed Affinity:
         The humanized antibody is guaranteed to have comparable binding affinity with parental antibody
  • Multiple Assays:
         Binding assays such as using ELISA , Biacore Surface Plasmon Resonance and a suite of biological assays are available

Customer Voice

“We are pleased with the antibody humanization service and the purification service of ProBio. It is very impressive and professional. we validated the humanization service by testing the antibody against anti-human IgG as the secondary antibody. The humanized antibody purification met our expectations and need. The characterization of the antibody that was highlighted by the COA is excellent. We would continue to avail your service and will also recommend it to anyone who needs such service. Thanks.” – Govind Panchamoorthy, Director, Genus Oncology

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