Antibody Humanization and Optimization Services

• Detailed sequence analysis and 3D structure modeling
• Identification of the best human germline sequence templates, grafting the rodent antibody or nanobody CDR regions into the human antibody frameworks
• Identify back-mutations and remove "hot spots" with developability issues
• High-throughput lead selection and evaluation
• Efficient lead preparation for research purpose in house (up to 20mg), CMC level protein preparation at the facility of our CMC partner

• One Fv 3D model developed using AlphaFold
• Up to 5 humanized VH and 5 humanized VL versions will be designed respectively
• PTMs will be removed if antibody`s affinity and/or developability won`t be significantly affected
• Change of 3D PI and aggregation potential will be marked during the design process, if developability issue is observed on bench, this information will be applied to resolve it
• Predicted high risk T cell epitope, B cell epitope, MHC-II epitope and antigenicity epitope will be recorded.

First round of leads selection and production

• DNA synthesis, cloning and transient transfection
• Affinity (ELISA) assay with expression media
• Aggregation potential and thermostability screening (ELISAs after heating) with expression media
• Production of top 3 humanization leads and chimeric antibody <2mg each)

Second round of final humanization lead selection (from first round leads)

• Affinity: Probelife (similar to Octect for detecting KD value) or cell-based assay (such as FACS, if other methods are not applicable)
• Aggregation potential: DLS
• Thermostability: DSF or DSC
• Purity: Non-reducing and reducing CE