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.

1. Study report, including sequences of the humanized antibodies. 2. All remaining purified antibodies.

Material Required:
1. Sequences of VL and VH. 2. Antigen and assay for measuring binding and affinity.

8 to 9 weeks