GenScript USA. FcB6.1 variants were codon optimized by using DNAworks with standard E. coli codons and amplified from a gBlock (IDT DNA)....">
Products/Services Used | Details | Operation |
---|
Computational design provides the opportunity to program protein-protein interactions for desired applications. We used de novo protein interface design to generate a pH-dependent Fc domain binding protein that buries immunoglobulin G (IgG) His-433. Using next-generation sequencing of naïve and selected pools of a library of design variants, we generated a molecular footprint of the designed binding surface, confirming the binding mode and guiding further optimization of the balance between affinity and pH sensitivity. In biolayer interferometry experiments, the optimized design binds IgG with a Kd of ∼4 nM at pH 8.2, and approximately 500-fold more weakly at pH 5.5. The protein is extremely stable, he... More