The human anti-mouse antibody (HAMA) reaction limits the therapeutic utility of mouse monoclonal antibodies. Humanization of mouse monoclonal antibodies by genetic engineering can improve the in-vivo tolerability by increasing their resemblance to antibodies naturally generated in humans.
Sino Biological delivers high-quality monoclonal antibody humanization services with a high degree of success (>90%) utilizing complementarity-determining region (CDR) grafting technology and computer-aided molecular modeling.
Image Credit: Sino Biological Inc.
Service highlights
- Humanization finalized within six days
- Humanized antibodies guaranteed to have a similar binding affinity to chimeric antibodies
- The 3D molecular modeling platform ensures the preservation of biological functions
- >90% human content of humanized antibodies
Service details for antibody humanization
Timeline: 6–8 weeks
Affinity validation
- Mouse antibody/antigen affinity validation
Preparation of human-mouse chimeric antibody
- Chimeric antibody–antigen interaction detection
- Transient expression and purification in HEK293 cells
Preparation of humanized variants
- CDR structural analysis
- SDS-PAGE and UV analysis
- 3D structural modeling of antibodies
- Affinity detection: ELISA, Biacore and Octet
- Transient transfection of HEK293 cells and purification
- Vector construction and plasmid preparation
Deliverables
- Antibody sequences
- Remaining antigens
- Top three humanized antibodies, 0.2~0.5 mg per strain (at least one humanized ab has an equivalent affinity with parental ab)
- CoA
- H and L chains in separate expression vectors
Customers provide:
- Protein antigens: purity >90%, concentration >1 mg/mL, and PBS formulation
- Sequences of the original mouse mAbs
Featured case study of antibody humanization service
As illustrated in the figure below, the affinity (KD) of the mouse antibody improved by over 38% after humanization.
Affinity of antibody before and after humanization. Image Credit: Sino Biological Inc.
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