Engineered pH-Sensitive Protein G/IgG Interaction

ACS Chem Biol. 2021 Jul 16;16(7):1142-1146. doi: 10.1021/acschembio.0c00943. Epub 2021 Jun 21.

Abstract

While natural protein-protein interactions have evolved to be induced by complex stimuli, rational design of interactions that can be switched-on-demand still remain challenging in the protein design world. Here, we demonstrate that a computationally redesigned natural interface for improved binding affinity could further be mutated to adopt a pH switchable interaction. The redesigned interface of Protein G/human IgG Fc domain (referred to as PrG/hIgG), when incorporated with histidine and glutamic acid on PrG (PrG-EHHE), showed a switch in binding affinity by 50-fold when the pH was altered from mild acidic to mild basic. The wild-type (WT) interface showed a negligible switch. The overall binding affinity under mild acidic pH for PrG-EHHE outperformed the wild-type PrG (PrG-WT) interaction. The new reagent PrG-EHHE can be revolutionary in IgG purification, since the standard method of using an extreme acidic pH for elution can be circumvented.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Bacterial Proteins / chemistry
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Glutamic Acid / chemistry
  • Histidine / chemistry
  • Humans
  • Hydrogen-Ion Concentration
  • Immunoglobulin Fc Fragments / chemistry
  • Immunoglobulin Fc Fragments / genetics
  • Immunoglobulin Fc Fragments / metabolism*
  • Immunoglobulin G / chemistry
  • Immunoglobulin G / metabolism*
  • Mutation
  • Protein Binding
  • Protein Domains
  • Protein Engineering
  • Streptococcus / chemistry

Substances

  • Bacterial Proteins
  • IgG Fc-binding protein, Streptococcus
  • Immunoglobulin Fc Fragments
  • Immunoglobulin G
  • Glutamic Acid
  • Histidine