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ITC Measurement for High-Affinity Aptamers Binding to Their Target Proteins

  • Ryo Amano
  • Tomohisa Furukawa
  • Taiichi SakamotoEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1964)

Abstract

Aptamers are nucleic acid molecules that bind to a target molecule with high affinity and specificity, which are generated by a process known as systematic evolution of ligands by exponential enrichment (SELEX). Because of their high affinity and specificity, aptamers were developed as therapeutic agents. Although aptamers are investigated as promising therapeutic agents, the mechanism of their high affinity and specificity is not clear. Therefore, structural and biophysical studies are important to know that. To date, ITC is increasingly being used to study the thermodynamic basis of aptamer–target protein interactions. Understanding the mechanism of aptamer binding would contribute to their development for therapeutic applications. In this chapter, we describe the protocol to study the thermodynamics of aptamer–protein interactions.

Key words

Aptamer Protein C value Competition Heat capacity change 

Notes

Acknowledgments

This study was supported by research grants of Innovative Area, Structural Cell Biology (Number 23121528), and the Strategic Research Foundation Grant-aided Project for Private Universities (Number S1101001) from the Ministry of Education, Sports, Culture, Science and Technology (MEXT) of Japan.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Ryo Amano
    • 1
  • Tomohisa Furukawa
    • 1
  • Taiichi Sakamoto
    • 1
    Email author
  1. 1.Faculty of Advanced Engineering, Department of Life ScienceChiba Institute of TechnologyNarashino-shiJapan

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