Systematic investigation of optimal aptamer immobilization for protein-microarray applications

Analytical Chemistry
Johanna-Gabriela WalterThomas Scheper

Abstract

Aptamers are short single-stranded DNA or RNA oligonucleotides that can bind to a wide range of target molecules with high affinity and specificity. As nucleic acids, aptamers can undergo denaturation, but the process is reversible. As a result of this stability and the possibility of automated selection of aptamers, these oligonucleotides are highly promising capture molecules in microarray formats. In this study, his-tagged proteins and an aptamer directed against the his-tag were chosen as a model system. Different factors affect the activity of aptamers immobilized on a solid support like a microarray surface. The orientation of the immobilized aptamer plays an important role in correct aptamer folding and, thus, in effective binding of the corresponding target. Other important parameters identified in this work are the microarrays' surface charge as well as the length of the spacer between aptamer and solid support. These parameters were investigated systematically, resulting in the development of an aptamer-based microarray for detection of his-tagged proteins. The general applicability of the developed immobilization strategy was demonstrated by utilization of three different aptamers.

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Citations

Jan 21, 2009·Analytical and Bioanalytical Chemistry·Changjiang YouJacob Piehler
Jul 11, 2013·Applied Microbiology and Biotechnology·Michael MeyerJohanna-Gabriela Walter
Dec 5, 2012·Biosensors & Bioelectronics·Lin-Chi ChenKonan Peck
Nov 12, 2013·Journal of Biotechnology·Yoshimitsu TakakuraYoshimitsu Kakuta
Aug 25, 2010·Journal of Nanobiotechnology·Johanna G WalterStephan Barcikowski
Feb 19, 2011·Expert Review of Proteomics·Harini ChandraSanjeeva Srivastava
Dec 4, 2015·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Lewis A FraserJulian A Tanner
Jan 18, 2016·Molecular Immunology·Roald Nezlin
Apr 5, 2016·Journal of Nucleic Acids·Jennifer A MartinNancy Kelley-Loughnane
May 4, 2011·Biotechnology and Bioengineering·Öznur KökpinarThomas Scheper
Sep 25, 2010·Wiley Interdisciplinary Reviews. Systems Biology and Medicine·Yifat Merbl, Marc W Kirschner
Sep 8, 2016·Microarrays·Rebecca JonczykFrank Stahl
Jan 31, 2017·Journal of Biotechnology·Katharina UrmannThomas Scheper
Nov 8, 2013·Scientific Reports·Hong Yan Liu, Xiaohu Gao
Mar 3, 2018·International Journal of Molecular Sciences·Regina Stoltenburg, Beate Strehlitz
Mar 25, 2011·Angewandte Chemie·Yamuna Krishnan, Friedrich C Simmel
Sep 17, 2019·The Analyst·Nasser A AlamraniRuchi Gupta
Oct 17, 2019·Biosensors·Tracy KurthJohanna-Gabriela Walter
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Apr 10, 2012·Talanta·Ekaterina S SinitsynaTatiana B Tennikova

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