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Catalog #24401 - Antibody Humanization and Affinity Measurement
Title:
Antibody Humanization and Affinity Measurement
Description:
Humanization and Affinity Measurement
Scope:
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.