Efficient isolation and elution of cellular proteins using aptamer-mediated protein precipitation assay

Biochem Biophys Res Commun. 2014 May 23;448(1):114-9. doi: 10.1016/j.bbrc.2014.04.086. Epub 2014 Apr 24.

Abstract

Protein precipitation is one of the most widely used methods for antigen detection and purification in biological research. We developed a reproducible aptamer-mediated magnetic protein precipitation method that is able to efficiently capture, purify and isolate the target proteins. We discovered DNA aptamers having individually high affinity and specificity against human epidermal growth factor receptor (EGFR) and human insulin receptor (INSR). Using aptamers and magnetic beads, we showed it is highly efficient technique to enrich endogenous proteins complex and is applicable to identify physiologically relevant protein-protein interactions with minimized nonspecific binding of proteins. The results presented here indicate that aptamers would be applicable as a useful and cost-effective tool to identify the presence of the particular target protein with their specific protein partners.

Keywords: Aptamer; Dextran sulfate; Isolation; Protein precipitation; SELEX.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, CD / isolation & purification
  • Aptamers, Nucleotide / metabolism
  • Cell Line, Tumor
  • Dextran Sulfate
  • ErbB Receptors / isolation & purification
  • Fractional Precipitation / methods*
  • Humans
  • Magnetics
  • Proteins / isolation & purification*
  • Receptor, Insulin / isolation & purification
  • SELEX Aptamer Technique / methods*

Substances

  • Antigens, CD
  • Aptamers, Nucleotide
  • Proteins
  • Dextran Sulfate
  • EGFR protein, human
  • ErbB Receptors
  • INSR protein, human
  • Receptor, Insulin