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Proc Natl Acad Sci U S A. 2014 Jan 14;111(2):675-80. doi: 10.1073/pnas.1313605111. Epub 2013 Dec 31.

Computational design of a pH-sensitive IgG binding protein.

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  • 1Department of Biochemistry and Howard Hughes Medical Institute, University of Washington, Seattle, WA 98195.


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, heat-resistant and highly expressed in bacteria, and allows pH-based control of binding for IgG affinity purification and diagnostic devices.


antibody purification; computational interface design; pH-sensitivity

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