Designed ankyrin repeat proteins (DARPins) from research to therapy

Methods Enzymol. 2012;503:101-34. doi: 10.1016/B978-0-12-396962-0.00005-7.

Abstract

Designed ankyrin repeat proteins (DARPins) have been developed into a robust and versatile scaffold for binding proteins. High-affinity binders are routinely selected by ribosome display and phage display. DARPins have entered clinical trials and have found numerous uses in research, due to their high stability and robust folding, allowing many new molecular formats. We summarize the DARPin properties and highlight some biomedical applications. Protocols are given for labeling with dyes and polyethylene glycol, for quantitatively measuring binding to cell surface receptors by kinetics and thermodynamics, and for exploiting new engineering opportunities from using "click chemistry" with nonnatural amino acids.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Affinity Labels / chemistry
  • Amino Acid Sequence
  • Animals
  • Ankyrin Repeat*
  • Biomedical Research
  • Carrier Proteins / chemistry*
  • Click Chemistry
  • Drug Design*
  • Escherichia coli / chemistry
  • Fluorescent Dyes / chemistry
  • Genetic Vectors / chemistry
  • Humans
  • Molecular Sequence Data
  • Peptide Library
  • Polyethylene Glycols / chemistry
  • Protein Binding
  • Protein Engineering / methods*
  • Protein Stability
  • Proteins / chemistry
  • Proteins / therapeutic use*
  • Receptors, Cell Surface / chemistry
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / isolation & purification
  • Ribosomes / chemistry
  • Thermodynamics
  • Titrimetry
  • Viruses / chemistry

Substances

  • Affinity Labels
  • Carrier Proteins
  • Fluorescent Dyes
  • Peptide Library
  • Proteins
  • Receptors, Cell Surface
  • Recombinant Fusion Proteins
  • Polyethylene Glycols