WuXi Biologics
Offering End-to-End Solutions
Background
Client requested production of CYP-X protein starting from gene sequence and generate crystals from purified protein for structure analysis.
Challenge
Solution/Results
Background
Client requested delivery of IgM with 95% purity (as determined by SDS-PAGE and HPLC analysis).
Challenge
Solution/Results
Background
Client requested the production of Kinase-Y with high purity.
Challenge
Solution/Results
Background
Challenge
No functional hits specific to isoform 2 were identified by hybridoma, even after multiple campaigns.
Solution/Results
By leveraing our in-house human naïve library we were able to identify functional hits by just one campaign. The specific number of campaigns and leads found were shown in the table.
Background
Use WuXi’s proprietary human naive antibody library to identify antibodies specific to each of the two splicing variants of antigen 1 named Ag 1a and Ag 1b.
Challenge
Solution/Results
Human naïve library was panned on the splicing variant b of the human antigen 1 (Ag 1b) in the presence of splicing variant a (Ag 1a) as a competitor. Following panning, rounds 2 and 3 outputs were screened by monoclonal phage ELISA. Round 3 output shows significant enrichment with Ag 1b specific clones with concurrent reduction of Ag 1a/Ag 1b cross-reactive clones. Sequence analysis of round 3 hits identified 6 unique clones, 2 of which also showed strong binding to cell-surface expressed Ag 1b (data not shown here).
Background
Client requested affinity maturation for their wild-type murine antibody and create a more “human” antibody in the process.
Challenge
Move antibody affinity from moderate binding levels (Kd = 1 nM) to equal or beat benchmark molecule with a double-digit picomolar Kd value.
Solution/Results
1.Parsimonious mutagenesis of the VHCDR3 region
2.Random mutagenesis of both the VH and VL chains
Aim
Production of a high quality GPCR protein in insect cells
Reconstitution of GPCR into nanodiscs
Key Challenges
Expression level of GPCR was very low <0.2 mg/L
The protein was unstable and its conformation was very flexible
It was hard to reconstitute GPCR into nanodiscs
Results
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