Surface-immobilized peptide aptamers as probe molecules for protein detection

Analytical Chemistry
Steven JohnsonChristoph Wälti

Abstract

We demonstrate the use of surface-immobilized, oriented peptide aptamers for the detection of specific target proteins from complex biological solutions. These peptide aptamers are target-specific peptides expressed within a protein scaffold engineered from the human protease inhibitor stefin A. The scaffold provides stability to the inserted peptides and increases their binding affinity owing to the resulting three-dimensional constraints. A unique cysteine residue was introduced into the protein scaffold to allow orientation-specific surface immobilization of the peptide aptamer and to ensure exposure of the binding site to the target solution. Using dual-polarization interferometry, we demonstrate a strong relationship between binding affinity and aptamer orientation and determine the affinity constant KD for the interaction between an oriented peptide aptamer ST(cys+)_(pep9) and the target protein CDK2. Further, we demonstrate the high selectivity of the peptide aptamer STM_(pep9) by exposing surface-immobilized ST(cys+)_(pep9) to a complex biological solution containing small concentrations of the target protein CDK2.

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Citations

Nov 1, 2013·Chemical Reviews·Kepa Ruiz-MirazoAndrés de la Escosura
Feb 25, 2010·Protein Engineering, Design & Selection : PEDS·Toni HoffmannPaul Ko Ferrigno
Dec 18, 2009·Journal of Molecular Recognition : JMR·Rebecca L Rich, David G Myszka
May 28, 2011·Protein Engineering, Design & Selection : PEDS·Lukas Kurt Josef StadlerPaul Ko Ferrigno
Mar 28, 2013·International Journal of Molecular Sciences·Andrea ConidiDanny Huylebroeck
Nov 29, 2014·The Analyst·M RainaC Wälti
Sep 15, 2016·Nature Communications·José Juan-ColásSteven D Johnson
Jun 29, 2011·Oncotarget·Renaud SeigneuricCarmen Garrido
Nov 14, 2019·Sensors·Thomas Allsop, Ron Neal
Jan 4, 2012·Angewandte Chemie·Marco MasciniSara Tombelli
Jul 2, 2016·Essays in Biochemistry·Paul Ko Ferrigno
May 30, 2008·Langmuir : the ACS Journal of Surfaces and Colloids·Siyoung Choi, William L Murphy
Feb 2, 2010·The Journal of Physical Chemistry. B·Li Qiang ChenCheng Zhi Huang
Apr 17, 2010·Analytical Chemistry·Pedro EstrelaPiero Migliorato
Aug 10, 2010·The Journal of Physical Chemistry. B·Wael SolimanKamaljit Kaur
Sep 30, 2014·The Journal of Physical Chemistry. B·Zunliang WangCharles L Brooks
Mar 28, 2012·Langmuir : the ACS Journal of Surfaces and Colloids·Steven JohnsonChristoph Wälti
Apr 18, 2020·ACS Applied Materials & Interfaces·Roozbeh Siavash MoakharSara Mahshid

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