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Antibody Humanization & PTM Service

Antibody Humanization & PTM Service

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OVERVIEW

Antibody humanization involves modifying mouse-derived antibodies through gene cloning and DNA recombination, replacing most of their amino acid sequences with human counterparts. This process retains the affinity and specificity of the original mouse monoclonal antibody while significantly reducing immunogenicity. Humanized antibodies are less likely to trigger immune responses in humans and are crucial for the development of therapeutic antibodies.

Gene Universal offers reliable antibody humanization services based on CDR grafting or surface resurfacing strategies. With an efficient workflow in place, we can complete humanization in as little as 3 weeks.The humanized antibodies retain the full affinity and specificity of their parental counterparts.

Post-translational modifications (PTMs) refer to chemical changes that occur to a protein after translation. During antibody production, storage, and clinical application, various PTM variants may arise. These modifications can alter the antibody’s charge and structure, potentially impacting its affinity for antigens and Fc receptors, and ultimately affecting critical quality attributes such as therapeutic activity. At Gene Universal, we use computational modeling combined with site-saturation mutagenesis libraries to predict and screen for mutants that retain affinity and specificity comparable to the original antibody—helping to streamline downstream therapeutic antibody development.

SERVICE ADVANTAGES

  • 01Antibody engineering guided by bioinformatics and structural biology through computer-aided design
  • 03Maintains affinity comparable to the original antibody
  • 02Simultaneous optimization of both heavy and light chains, achieving >90% humanization
  • 04Humanized antibody delivery in as fast as 3 weeks

SERVICE PROCEDURE

  • Start >
  • Week 1 >
  • Week 2 >
  • Week 3 >
  • Week 4 >
  • Week 5 >
  • Sequence Analysis & Homology Modeling

    Computational modeling / Germline selection

  • Sequence Design & Vector Construction

    Back-mutation / PTM consideration / Site-directed mutation library

  • Transient Expression & Purification

    96-well high-throughput plate screening / Small-scale expression(3–30 mL)and purification

  • Affinity Assessment

    BLI-based high-throughput affinity ranking / Biacore affinity validation

  • Antibody Delivery & Biacore Validation

    SDS-PAGE / SEC-HPLC / Biacore confirmation of affinity

SERVICE CONTENT

Items Service Details EDT Deliverable
Sequence Analysis

Construct 3D antibody structure models

Germline Selction

CDR grafting with key residue back mutations

Removal of PTM sites (optional)

1 Week

2–3 humanized antibody variants(at least one with affinity comparable to the original antibody)

Cloned plasmids

Humanization design report

2–3 antibody samples(1 mg each)

COA

Antibody Expression

Gene synthesis & vector construction

High-throughput expression in mammalian system or small-scale antibody expression

2 Week
Affinity Measurement

ELISA/BLI-based affinity screening from culture supernatant

Biacore validation

1 Week
Antibody Expression

Small-scale antibody expression and purification

QC

1 Week

CASE STUDY

parent-antibody
humanized-ab
Name Capture Level(RU) Analyte ka(1/Ms) kd(1/s) KD(M) Rmax(RU) Chi²(RU²) U-value
Parent Ab 102 XXX 8.06E+05 1.36E-04 1.68E-10 60.51 0.0288 2
Humanized Ab 156 XXX 4.28E+05 6.04E-05 1.41E-10 78.85 0.035 5

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