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Analytics & Formulation
- Purity Analysis by CE-SDS
- Aggregation Analysis
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- De Novo Antibody Sequencing
- Intact Mass Analysis
- Sequence Confirmation by Mass Spectrometry
- N-terminal Sequencing
- Post-Translational Modification
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Catalog #24401 - Antibody Humanization and Affinity Measurement
Antibody Humanization and Affinity Measurement
Humanization and Affinity Measurement
1. Antibody sequence analysis and homology modeling of mAb 3D structure.
2. Identification of key positions supporting CDR loop structure and VH-VL interface.
3. Design humanized variants (3VH, 3VL).
4. Assess the humanness of humanized variants by T20 humanness score developed at LakePharma. [Sean H Gao, Kexin Huang, Hua Tu, and Adam S Adler. (2013) Monoclonal antibody humanness score and its applications. BMC Biotechnology, 13:55]
5. Construct and express nine humanized antibodies (each combination of three designed heavy chains and three designed light chains) as well as a chimeric version (rodent variable regions, human constant regions) at the 10 mL scale in CHO cells (TunaCHO™ Process).
6. Assess expression levels and measure affinity by Octet, or competitive binding and compare to chimera.