Details
| Original language | English |
|---|---|
| Pages (from-to) | 7372-7378 |
| Number of pages | 7 |
| Journal | Analytical chemistry |
| Volume | 80 |
| Issue number | 19 |
| Publication status | Published - 27 Aug 2008 |
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.
ASJC Scopus subject areas
- Chemistry(all)
- Analytical Chemistry
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In: Analytical chemistry, Vol. 80, No. 19, 27.08.2008, p. 7372-7378.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Systematic Investigation of Optimal Aptamer Immobilization for Protein-Microarray Applications
AU - Walter, Johanna Gabriela
AU - Kökpinar, Öznur
AU - Friehs, Karl
AU - Stahl, Frank
AU - Scheper, Thomas
PY - 2008/8/27
Y1 - 2008/8/27
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=54749156725&partnerID=8YFLogxK
U2 - 10.1021/ac801081v
DO - 10.1021/ac801081v
M3 - Article
C2 - 18729475
AN - SCOPUS:54749156725
VL - 80
SP - 7372
EP - 7378
JO - Analytical chemistry
JF - Analytical chemistry
SN - 0003-2700
IS - 19
ER -