Abstract
The ability to monitor biomolecular recognition such as DNA hybridization and enzymatic reactivity in solutions with high sensitivity is important for developing effective bioassay strategies. Surface enhanced Raman scattering (SERS) based on use of solid substrates to produce the SERS effect for the detection often requires substrate preparation which is ineffective for rapid monitoring. This report describes a new strategy exploiting a gold nanoparticle (AuNP) based interparticle "hot-spot" for SERS monitoring of DNA mediated assembly and enzyme induced cleavage of the assembly in solution phase. The DNAs consist of two different complementary DNA strands with a thiol modification for attachment to AuNPs of selected sizes. In a solution containing AuNPs conjugated with one of the single-stranded (ss) DNA and other AuNPs labeled with a Raman reporter molecule, 4-mercaptobenzoic acid (MBA), the introduction of the complementary DNA strand leads to a linkage of the two types of AuNPs, producing double-stranded (ds) DNA-AuNP assembly (ds-DNA-AuNPs) with an interparticle "hot-spot" for SERS detection of the diagnostic bands of the reporter. Upon introducing a restriction enzyme (e.g. MspI) into the ds-DNA-AuNP assembly solution, the removal of the interparticle "hot-spot" due to restriction enzyme cleavage of the ds-DNA leads to a decrease of the SERS signals. While the detailed cleavage process may depend on the reaction time and the amount of enzyme, the viability of using gold nanoparticle "hot-spot" based SERS monitoring of DNA assembly and enzyme cleavage is clearly demonstrated, which has important implications for developing new strategies for bioassays.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 4941-4949 |
| Number of pages | 9 |
| Journal | Analyst |
| Volume | 138 |
| Issue number | 17 |
| DOIs | |
| State | Published - Jan 1 2013 |
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All Science Journal Classification (ASJC) codes
- Analytical Chemistry
- Biochemistry
- Environmental Chemistry
- Spectroscopy
- Electrochemistry
Cite this
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DNA assembly and enzymatic cutting in solutions : A gold nanoparticle based SERS detection strategy. / Crew, Elizabeth; Yan, Hong; Lin, Liqin; Yin, Jun; Skeete, Zakiya; Kotlyar, Timur; Tchah, Nuri; Lee, Jehwan; Bellavia, Michael; Goodshaw, Isaac; Joseph, Pharrah; Luo, Jin; Gal, Susannah; Zhong, Chuan Jian.
In: Analyst, Vol. 138, No. 17, 01.01.2013, p. 4941-4949.Research output: Contribution to journal › Article
TY - JOUR
T1 - DNA assembly and enzymatic cutting in solutions
T2 - A gold nanoparticle based SERS detection strategy
AU - Crew, Elizabeth
AU - Yan, Hong
AU - Lin, Liqin
AU - Yin, Jun
AU - Skeete, Zakiya
AU - Kotlyar, Timur
AU - Tchah, Nuri
AU - Lee, Jehwan
AU - Bellavia, Michael
AU - Goodshaw, Isaac
AU - Joseph, Pharrah
AU - Luo, Jin
AU - Gal, Susannah
AU - Zhong, Chuan Jian
PY - 2013/1/1
Y1 - 2013/1/1
N2 - The ability to monitor biomolecular recognition such as DNA hybridization and enzymatic reactivity in solutions with high sensitivity is important for developing effective bioassay strategies. Surface enhanced Raman scattering (SERS) based on use of solid substrates to produce the SERS effect for the detection often requires substrate preparation which is ineffective for rapid monitoring. This report describes a new strategy exploiting a gold nanoparticle (AuNP) based interparticle "hot-spot" for SERS monitoring of DNA mediated assembly and enzyme induced cleavage of the assembly in solution phase. The DNAs consist of two different complementary DNA strands with a thiol modification for attachment to AuNPs of selected sizes. In a solution containing AuNPs conjugated with one of the single-stranded (ss) DNA and other AuNPs labeled with a Raman reporter molecule, 4-mercaptobenzoic acid (MBA), the introduction of the complementary DNA strand leads to a linkage of the two types of AuNPs, producing double-stranded (ds) DNA-AuNP assembly (ds-DNA-AuNPs) with an interparticle "hot-spot" for SERS detection of the diagnostic bands of the reporter. Upon introducing a restriction enzyme (e.g. MspI) into the ds-DNA-AuNP assembly solution, the removal of the interparticle "hot-spot" due to restriction enzyme cleavage of the ds-DNA leads to a decrease of the SERS signals. While the detailed cleavage process may depend on the reaction time and the amount of enzyme, the viability of using gold nanoparticle "hot-spot" based SERS monitoring of DNA assembly and enzyme cleavage is clearly demonstrated, which has important implications for developing new strategies for bioassays.
AB - The ability to monitor biomolecular recognition such as DNA hybridization and enzymatic reactivity in solutions with high sensitivity is important for developing effective bioassay strategies. Surface enhanced Raman scattering (SERS) based on use of solid substrates to produce the SERS effect for the detection often requires substrate preparation which is ineffective for rapid monitoring. This report describes a new strategy exploiting a gold nanoparticle (AuNP) based interparticle "hot-spot" for SERS monitoring of DNA mediated assembly and enzyme induced cleavage of the assembly in solution phase. The DNAs consist of two different complementary DNA strands with a thiol modification for attachment to AuNPs of selected sizes. In a solution containing AuNPs conjugated with one of the single-stranded (ss) DNA and other AuNPs labeled with a Raman reporter molecule, 4-mercaptobenzoic acid (MBA), the introduction of the complementary DNA strand leads to a linkage of the two types of AuNPs, producing double-stranded (ds) DNA-AuNP assembly (ds-DNA-AuNPs) with an interparticle "hot-spot" for SERS detection of the diagnostic bands of the reporter. Upon introducing a restriction enzyme (e.g. MspI) into the ds-DNA-AuNP assembly solution, the removal of the interparticle "hot-spot" due to restriction enzyme cleavage of the ds-DNA leads to a decrease of the SERS signals. While the detailed cleavage process may depend on the reaction time and the amount of enzyme, the viability of using gold nanoparticle "hot-spot" based SERS monitoring of DNA assembly and enzyme cleavage is clearly demonstrated, which has important implications for developing new strategies for bioassays.
UR - http://www.scopus.com/inward/record.url?scp=84881192427&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84881192427&partnerID=8YFLogxK
U2 - 10.1039/c3an00683b
DO - 10.1039/c3an00683b
M3 - Article
C2 - 23799231
AN - SCOPUS:84881192427
VL - 138
SP - 4941
EP - 4949
JO - The Analyst
JF - The Analyst
SN - 0003-2654
IS - 17
ER -