Immunoassays based on directional surface plasmon-coupled emission

Matveeva, Evgenia G.; Gryczyński, Zygmunt; Gryczyński, Ignacy; Lakowicz, Joseph R.

doi:10.1016/j.jim.2003.12.009

Cited by 40 publications

(35 citation statements)

References 23 publications

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“…The enhanced excitation field is confined to the evanescent layer, effectively reducing the background from the sample volume matrix. The exceptional sensitivity of SPCE-based assays has been observed in our and other laboratories (42)(43)(44)(45)(46)(47)(48).…”

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confidence: 55%

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Plasmonic Technology: Novel Approach to Ultrasensitive Immunoassays

Lakowicz¹,

Malicka²,

Matveeva³

et al. 2005

Clinical Chemistry

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, we have taken advantage of the favorable properties of surface plasmon-coupled emission (SPCE) to improve fluorescence-based immunoassays. SPCE occurs when excited fluorophores near conducting metallic structures efficiently couple to surface plasmons. These surface plasmons, appearing as free electron oscillations in the metallic layer, produce electromagnetic radiation that preserves the spectral properties of fluorophores but is highly polarized and directional. SPCE immunoassays provide several advantages over other fluorescencebased methods. This review explains new approaches to fluorescence immunoassays, including our own use of SPCE for simultaneous detection of more than one fluorescent marker and performance of immunoassays in the presence of an optically dense medium, such as whole blood. © 2005 American Association for Clinical ChemistryBiological markers in physiologic fluids are very useful tools in clinical diagnostics. Sensitive and reliable detection of specific biomarkers is crucial in disease identification, therapy, and patient screening. Reliable and quick detection of low concentrations of markers is particularly important in a disease such as acute myocardial infarction (AMI).1 Myoglobin (Myo), although not completely cardiac specific, is one of the earliest markers to increase after AMI (1-6 ) and has been recommended for use in combination with other markers, such as creatine kinase-MB, troponin I, and troponin T, as an early diagnostic indicator for AMI (1,(7)(8)(9)(10). The immunoassay technique is widely used in this procedure (11 ).Immunoassay based on fluorescence detection is one approach to high-sensitivity detection of biomarkers (12)(13)(14)(15)(16) . Different fluorescence detection approaches include polarization (17-21 ), resonance energy transfer (22-24 ), and time-resolved gated assays based on long-lived lanthanide emission (25-28 ). New approaches to fluorescence immunoassays are being developed, including multiphoton excitation (29 -31 ), with the emphasis on highthroughput immunoassays (32-35 ).The sensitivity of fluoroimmunoassays is typically limited by background fluorescence, which is present in most biological samples and in the optical elements of the instrumentation. In this report, we describe a new immunoassay format that provides increased sensitivity and background rejection by efficient light collection of emissions occurring near the bioaffinity surface. In this approach, a fluorescently labeled reporting molecule (antibody) is bound near a metallic surface, and the binding produces a highly directional and polarized emission. This effect is based on the resonant coupling of excited fluorophores with collective electron motions/oscillations at the interface between a thin metal film (typically silver or gold) and a dielectric bulk material. The so-called surface plasmons comprise an electromagnetic wave confined to the interface between the metal film and the transparent medium, and the electromagnetic wave of the surface plasmons is coupled to oscillatio...

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confidence: 55%

“…We used SPCE to develop a model affinity assay with labeled goat anti-rabbit IgG antibodies against rabbit IgG bound to a 50-nm thick silver film (48 ). Rhodamine Red-X-labeled IgG was placed near the silver surface by its binding to the surface-bound antigen (Fig.…”

Section: Model Spce Immunoassaymentioning

confidence: 99%

Plasmonic Technology: Novel Approach to Ultrasensitive Immunoassays

Lakowicz¹,

Malicka²,

Matveeva³

et al. 2005

Clinical Chemistry

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“…How can SBCS monitor the cellular effects? As is well known, SPR biosensors could detect the changes of molecular concentration in the range of the evanescent wave, which is within a few hundred nm from the sensor surface [13] . Diameter of C6 cell is tenths of microns and the cell membrane is only 7.5-10 nm wide, the measured signal could only be the reflection of the regular redistribution of intracellular components near the cell membrane.…”

Section: Discussionmentioning

confidence: 99%

Using of the surface plasmon resonance cytosensor for real-time and non-invasive monitoring of cellular effects in living C6 cells induced by PMA

Wang

Xiong

et al. 2006

CHINESE SCI BULL

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Developing novel instruments and technologies for spatio-temporal and dynamic measurements of the intricate cellular effects involving molecular translocation, signal transduction, and molecular interactions inside living cells is essential for the cell and molecular biology science. For the purpose of monitoring and investigating molecular events in living cells at real-time, the surface plasmon resonance based cytosensor (SBCS) for cell culturing and signal monitoring was established, and on the basis of it, the corresponding technology was also established by monitoring and analyzing SPR responses induced in rat C6 glioma cells by phorbol 12-myristate 13-acetate (PMA). The SPR signals induced by PMA in living C6 cells were significantly different from those groups without cells. These responses were strongly dependent on and saturable to the concentrations of PMA, and could be suppressed by the specific and potent PKC inhibitors, which indicated that the measured signal could be the reflection of the redistribution of intracellular components near the cell membrane triggered by the activation of PKC. This research provides a quantitative and non-invasive technique to study the spatio-temporal characteristics of the cellular effects in living cells at real-time. Furthermore, this technology could also be widely used in the basic research as well as applied realms, such as space effects evaluation, environmental safety assessment, biological weapon detection, cellular and molecular research, and drug screening.

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“…We tested two kinds of model background solution, a highly fluorescent (non-binding labeled antibodies) and a highly absorbing (17% hemoglobin solution). We found that the use of SPCE provides very effective background suppression: the emission from labeled unbound antibodies was suppressed over 100-fold in the KR (versus RK) configuration, and the highly absorbing hemoglobin solution reduced the detected SPCE fluorescence intensity only about 50% [12]. We also tested human serum and human whole blood matrix [15].…”

Section: Introductionmentioning

confidence: 97%

“…Recently, we introduced a model immunoassay [12][13] and a myoglobin immunoassay [14] on a thin silver metal surface utilizing SPCE. We tested two kinds of model background solution, a highly fluorescent (non-binding labeled antibodies) and a highly absorbing (17% hemoglobin solution).…”

Section: Introductionmentioning

confidence: 99%

Ultrasensitive detection in optically dense physiological media: applications to fast reliable biological assays

et al. 2006

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We present a novel approach for performing fluorescence immunoassay in serum and whole blood using fluorescently labeled anti-rabbit IgG. This approach, which is based on Surface Plasmon-Coupled Emission (SPCE), provides increased sensitivity and substantial background reduction due to exclusive selection of the signal from the fluorophores located near a bio-affinity surface. Effective coupling range for SPCE is only couple of hundred nanometers from the metallic surface. Excited fluorophores outside the coupling layer do not contribute to SPCE, and their free-space emission is not transmitted through the opaque metallic film into the glass substrate. An antigen (rabbit IgG) was adsorbed to a slide covered with a thin silver metal layer, and the SPCE signal from the fluorophore-labeled anti-rabbit antibody, binding to the immobilized antigen, was detected. The effect of the sample matrix (buffer, human serum, or human whole blood) on the end-point immunoassay SPCE signal is discussed. The kinetics of binding could be monitored directly in whole blood or serum. The results showed that human serum and human whole blood attenuate the SPCE end-point signal and the immunoassay kinetic signal only approximately 2-and 3-fold, respectively (compared to buffer), resulting in signals that are easily detectable even in whole blood. The high optical absorption of the hemoglobin can be tolerated because only fluorophores within a couple of hundred nanometers from the metallic Downloaded From: http://proceedings.spiedigitallibrary.org/ on 06/15/2016 Terms of Use: http://spiedigitallibrary.org/ss/TermsOfUse.aspx 635 nm Excitation / / / N C pfilm contribute to SPCE. Both glass and plastic slides can be used for SPCE-based assays. We believe that SPCE has the potential of becoming a powerful approach for performing immunoassays based on surface-bound analytes or antibodies for many biomarkers directly in dense samples such as whole blood, without any need for washing steps.

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Immunoassays based on directional surface plasmon-coupled emission

Cited by 40 publications

References 23 publications

Plasmonic Technology: Novel Approach to Ultrasensitive Immunoassays

Plasmonic Technology: Novel Approach to Ultrasensitive Immunoassays

Using of the surface plasmon resonance cytosensor for real-time and non-invasive monitoring of cellular effects in living C6 cells induced by PMA

Ultrasensitive detection in optically dense physiological media: applications to fast reliable biological assays

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