Isolation of Artificial Binding Proteins (Affimer Reagents) for Use in Molecular and Cellular Biology

Methods Mol Biol. 2021:2247:105-121. doi: 10.1007/978-1-0716-1126-5_6.

Abstract

Artificial binding proteins have been validated as alternatives to antibodies in their use as research reagents in molecular and cellular biology. For example, they have been used as inhibitors of protein-protein interactions to modulate activity, to facilitate crystallization, and as probes for cellular imaging.Phage display is a widely used approach for isolating target-specific binding reagents, and it has even been used to isolate isoform-specific binding proteins and binders that can distinguish between highly homologous protein domains.Here, we describe methods that have been employed in isolating highly specific artificial binding proteins against a wide range of target proteins.

Keywords: Affimer; Artificial binding protein; Inhibitor; Panning; Phage ELISA; Phage display; Protein–protein interaction; Research reagents.

MeSH terms

  • Antibodies / metabolism
  • Carrier Proteins / chemistry
  • Carrier Proteins / isolation & purification*
  • Cell Biology*
  • Cell Surface Display Techniques
  • Cytological Techniques
  • Enzyme-Linked Immunosorbent Assay
  • Humans
  • Indicators and Reagents*
  • Molecular Biology* / methods
  • Peptide Library
  • Protein Binding
  • Structure-Activity Relationship

Substances

  • Antibodies
  • Carrier Proteins
  • Indicators and Reagents
  • Peptide Library