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Enrichment and detection of rare proteins with aptamer-conjugated gold nanorods.

ABSTRACT: Rare protein enrichment and sensitive detection hold great potential in biomedical studies and clinical practice. This work describes the use of aptamer-conjugated gold nanorods for the efficient enrichment of rare proteins from buffer solutions and human plasma. Gold nanorod (AuNR) surfaces were modified with a long PEG chain and a 15-mer thrombin aptamer for protein enrichment and detection. Studies of the effect of surface modification on enrichment efficiency of thrombin showed that a change of only one EG(6) linker unit, i.e., from 2EG(6) to 3EG(6), could increase thrombin protein capture efficiency by up to 47%. Furthermore, a 1 ppm sample of thrombin in buffer could be enriched with around 90% efficiency using a low concentration (0.19 nM) of gold nanorod probe modified with 3EG(6) spacer, and with the same probe, effective capture was achieved down to 10 ppb (1 ng) thrombin in plasma samples. In addition to ?-thrombin enrichment, prothrombin was also efficiently captured from plasma samples via gold nanorods conjugated with 15-mer thrombin aptamer. Our work demonstrates efficient enrichment of rare proteins using aptamer-modified nanomaterials, which can be used in biomarker discovery studies.

SUBMITTER: Yasun E

PROVIDER: S-EPMC3407574 | biostudies-literature | 2012 Jul

REPOSITORIES: biostudies-literature

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Enrichment and detection of rare proteins with aptamer-conjugated gold nanorods.

Yasun Emir E Gulbakan Basri B Ocsoy Ismail I Yuan Quan Q Shukoor Mohammed Ibrahim MI Li Chunmei C Tan Weihong W

Analytical chemistry 20120625 14

Rare protein enrichment and sensitive detection hold great potential in biomedical studies and clinical practice. This work describes the use of aptamer-conjugated gold nanorods for the efficient enrichment of rare proteins from buffer solutions and human plasma. Gold nanorod (AuNR) surfaces were modified with a long PEG chain and a 15-mer thrombin aptamer for protein enrichment and detection. Studies of the effect of surface modification on enrichment efficiency of thrombin showed that a change ...[more]

PMID: 22725611

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Project description:Abscisic acid (ABA) is a plant hormone, which plays an important role in plant growth, crop cultivation and modern agricultural engineering management. Accordingly, the detection of ABA content combined with new techniques and methods has become a more and more popular problem in the field of agricultural engineering. In this work, a SERRS and fluorescence dual-function sensor based on the fluorescence quenching and Raman enhancement properties of gold nanorods (AuNRs) was developed, and applied to the detection of plant hormone ABA. The dual-function reporter molecule Rhodamine isothiocyanate (RBITC) and complementary DNA (cDNA) were modified on AuNRs (AuNRs@RBITC@cDNA) as signal probes and aptamer modified magnetic nanoparticles (Fe3O4MNPs@Apt) as capture probes. Through the specific recognition of ABA aptamer and its complementary chains, an dual-function aptamer sensor based on SERRS and fluorescence was constructed. When ABA molecules were present in the detection system, the signal probes were detached from the capture probes due to the preferential binding between aptamer and ABA molecules. SERS signal of the reporter molecules appeared in the supernatant after magnetic separation, and it increased with the increase of ABA concentration. If the etching agent that can etch AuNRs was added to the supernatant, the AuNRs was etching disappeared, then the signal molecules fall off from the AuNRs, and the fluorescence signal intensity would recovered. The intensity of fluorescence signal also increased with the increase of ABA concentration. Thus, the quantitative relationship between ABA concentration and SERRS intensity and fluorescence intensity of signal molecules was established. The linear range of SERRS detection was 100 fM–0.1 nM, the detection limit was 38 fM; The linear range of fluorescence detection was 1 pM–100 nM, the detection limit is 0.33 p.m. The constructed dual-effect sensor was used in the recovery laboratory of real ABA samples, the recovery rate was up to 85–108%.

Dataset Information

Enrichment and detection of rare proteins with aptamer-conjugated gold nanorods.

Publications

Enrichment and detection of rare proteins with aptamer-conjugated gold nanorods.

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