Single-Molecule Surface-Enhanced Raman and Fluorescence Correlation Spectroscopy of Horseradish Peroxidase

Bjerneld, Erik J.; Földes-Papp, Zeno; Käll, Mikael; Rigler, Rudolf

doi:10.1021/jp012268y

Cited by 127 publications

(106 citation statements)

References 36 publications

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“…Several examples of applications of SM-SERS have been proposed since its discovery. [26][27][28][29] These include the detection and characterization of proteins, [30][31][32][33] the investigation of surface dynamics, 34 and even DNA sequencing. 28 However, there are no reports to date on the application of SM-SERS to study processes in an electrochemical environment.…”

Section: Introductionmentioning

confidence: 99%

Electrochemical Control of the Time-Dependent Intensity Fluctuations in Surface-Enhanced Raman Scattering (SERS)

et al. 2009

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Time-dependent fluctuations in surface-enhanced Raman scattering (SERS) intensities were recorded from a roughened silver electrode immersed in diluted solutions of rhodamine 6G (R6G) and congo red (CR). These fluctuations were attributed to a small number of SERS-active molecules probing regions of extremely high electromagnetic field (hot spots) at the nanostructured surface. The time-dependent distribution of SERS intensities followed a tailed statistics at certain applied potentials, which has been linked to single-molecule dynamics. The shape of the distribution was reversibly tuned by the applied voltage. Mixtures of both dyes, R6G and CR, at low concentrations were also investigated. Since R6G is a cationic dye and CR is an anionic dye, the statistics of the SERS intensity distribution of either dye in a mixture were independently controlled by adjusting the applied potential. The potential-controlled distribution of SERS intensities was interpreted by considering the modulation of the surface coverage of the adsorbed dye by the interfacial electric field. This interpretation was supported by a two-dimensional Monte Carlo simulation that took into account the time evolution of the surface configuration of the adsorbed species and their probability to populate a hypothetical hot spot. The potential-controlled SERS dynamics reported here is a first step toward the spectroelectrochemical investigation of redox processes at the single-molecule level by SERS.

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Section: Introductionmentioning

confidence: 99%

Electrochemical Control of the Time-Dependent Intensity Fluctuations in Surface-Enhanced Raman Scattering (SERS)

et al. 2009

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“…Upon exploiting such hot spots and the high sensitivity of modern Raman spectrometers, it is possible to achieve single molecule detection (SMD) using SERS or surface-enhanced resonance Raman scattering (SERRS), which is the ultimate limit in terms of chemical analysis [20][21][22][23] . Since the highest enhancement factors are obtained with aggregates or clusters of nanoparticles (where hot spots are generated), there is a hefty number studies on the relationship of packing, size and shape of nanoparticles and the SERS signal 19,[24][25][26][27][28] . The reproducible fabrication of nanostructures with maximum enhancement factor, as required for SMD, remains a challenge though.…”

Section: Introductionmentioning

confidence: 99%

Surface-Enhanced Raman Scattering: Metal Nanostructures Coated With Langmuir-Blodgett Films

Aléssio

Constantino

Aroca

et al. 2010

J. Chil. Chem. Soc.

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on leave from the department of chem. and biochem, university of windsor, windsor, ontario, canada. (Received: August 30, 2010 -Accepted: November 16, 2010) aBStract This review deals with surface-enhancved Raman scattering (SERS) employing Langmuir-Blodgett (LB) films, which serve as model systems for developing theoretical and experimental studies to elucidate the SERS effect. In addition, LB films have be used as integral parts of molecular architectures for SERS-active substrates. On the other hand, SERS and surface-enhaced resonant Raman scattering (SERRS) have allowed various properties of LB films to be investigated, especially those associated with molecular-level interactions. In the paper, emphasis is placed on single molecule detection (SMD), where the target molecule is diluted on an LB matrix of spectral silent material (low Raman cross section). The perspectives and challenges for combining SERS and LB films are also discussed.

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“…Signal analysis of SERS was used to identify the native constituents of live epithelial cells employing endocytosed 60 nm gold nanoparticles [11]. SERS has also been used to differentiate bacteria from bacteriophages by conjugating them to 60nm gold nanoparticles [12] and in single molecule detection [13,14]. …”

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confidence: 99%

Identification of binding interactions between myeloperoxidase and its antibody using SERS

et al. 2010

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Surface Enhanced Raman Spectroscopy (SERS) is a widely used spectroscopic method that can dramatically increase the sensitivity of Raman spectroscopy and has demonstrated significant benefit in the identification of biological molecules. We report the use of SERS in differentiating the bound immunocomplex of myeloperoxidase (MPO) and its antibody from the unbound complex and its individual components. The SERS signal was enabled by gold nanoparticles attached to MPO, pAb and their immunocomplex at an excitation wavelength of 785 nm. The obtained SERS spectrum of MPO is in agreement with previous literature. Comparative SERS spectrum analysis of MPO, pAb, and their immunocomplex reveals the significant peak shifts and intensity variations caused by the conformational changes due to the immunocomplex formation. Several key areas have been identified which correspond to specific amino acids being shielded from undergoing resonance while new amino acid residues are made visible in the SERS spectrum of the immunocomplex and could be a result of conformational binding. Our work demonstrates the capability of SERS to identify binding events and differentiate an immunocomplex from its unbound components with direct applications in biosensors.Keywords: Gold nanoparticle; Antigen; Immunocomplex; Biosensors Citation: E. S. Papazoglou, S. Babu, S. Mohapatra, D. R. Hansberry and C. Patel, "Identification of binding interactions between myeloperoxidase and its antibody using SERS", Nano-Micro Lett. 2, 74-81 (2010). doi:10.5101/nml.v2i2. p74-82 Raman spectroscopy is a useful technique, based on Raman scattering, that has been widely used for molecular material EXPERIMENTAL METHODSAll chemicals were purchased from Sigma Aldrich unless otherwise mentioned, pAb (rabbit anti human myeloperoxidase)was purchased from ABD-Serotec and myeloperoxidase (MPO) was purchased from Lee Biosolutions Inc.Gold nanoparticles were prepared according to Frens, G.[24] with added modifications. 500µl of 1% chloroauric acid Slides with immobilized gold nanoparticles were then washed with DI water and air dried. A well with a capacity of ~100 mL was constructed using plastic pipette and epoxy on top of the glass slide to facilitate pAb immobilization and subsequent SERS data collection. -Micro Lett. 2, 74-82 (2010) Polyclonal antibody was immobilized by letting ~100 ml of 100 nM pAb solutions interact with the gold nanoparticle coated slides for 15 minutes at room temperature followed by thorough washing with 1x PBS buffer (pH7.4). Gold nanoparticles coated with pAb were then allowed to interact with MPO (100 μl, 1 μM) Nanofor 15 minutes at room temperature followed by washing. A similar procedure was followed to immobilize MPO (100 μl, 1 M) directly on gold nanoparticles. SERS data was collected from the Au-pAb conjugates, Au-pAb/MPO immunocomplex, and Au-MPO conjugates immediately after preparation using a Renishaw RM1000 confocal Raman microspectrometer with a 50x long focal microscope. A 785nm diode laser (15 mW) was used to collect the ...

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Single-Molecule Surface-Enhanced Raman and Fluorescence Correlation Spectroscopy of Horseradish Peroxidase

Cited by 127 publications

References 36 publications

Electrochemical Control of the Time-Dependent Intensity Fluctuations in Surface-Enhanced Raman Scattering (SERS)

Electrochemical Control of the Time-Dependent Intensity Fluctuations in Surface-Enhanced Raman Scattering (SERS)

Surface-Enhanced Raman Scattering: Metal Nanostructures Coated With Langmuir-Blodgett Films

Identification of binding interactions between myeloperoxidase and its antibody using SERS

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