Acoustic Immunosensing of Exosomes Using a Quartz Crystal Microbalance with Dissipation Monitoring

Exosomes are endocytic lipid-membrane bound bodies with potential to be used as biomarkers in cancer and neurodegenerative disease. The limitations and scarcity of current exosome characterisation approaches has led to a growing demand for translational techniques, capable of determining their molecular composition and physical properties in physiological fluids. Here, we investigate label-free immunosensing, using a quartz crystal microbalance with dissipation (QCM-D), to detect exosomes by exploiting their surface protein profile. Exosomes expressing the transmembrane protein CD63 were isolated by size-exclusion chromatography from cell culture media. QCM-D sensors functionalised with anti-CD63 antibodies formed a direct immunoassay towards CD63-positive exosomes, exhibiting a limit-of-detection of 1.7x10^8 and 1.1x10^8 exosome sized particles (ESPs) ml^-1 for frequency and dissipation response respectively, i.e., clinically relevant concentrations. Our proof-of-concept findings support the adoption of dual-mode acoustic analysis of exosomes, leveraging both frequency and dissipation monitoring for use in diagnostic assays.

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Immunoassay of Glomalin by Quartz Crystal Microbalance Biosensor Containing Iron Oxide Nanoparticles

International Journal of Analytical Chemistry ◽

10.1155/2020/8844151 ◽

2020 ◽

Vol 2020 ◽

pp. 1-6

Author(s):

Miroslav Pohanka ◽

Vitezslav Vlcek

Keyword(s):

Iron Oxide ◽

Iron Oxide Nanoparticles ◽

Quartz Crystal Microbalance ◽

Quartz Crystal ◽

Mechanical Stability ◽

Limit Of Detection ◽

Oxide Nanoparticles ◽

Label Free ◽

Protein Determination ◽

Bradford Assay

Glomalin is a soil protein resembling heat shock protein (HSP) 60 and exerting high affinity to metals, causing retention of water in the environment and improving mechanical stability of soil. Currently, glomalin is determined in the soil or other samples by combination of autoclaving extraction and total protein determination typically by the Bradford method. In this paper, a piezoelectric biosensor was prepared to determine glomalin in a label-free measurement. The biosensor contained antibodies immobilized on quartz crystal microbalance (QCM), and the recognition layer was stabilized by iron oxide nanoparticles. The assay was tested on real soil samples and compared with the standard Bradford assay. Limit of detection of the assay was equal to 2.4 µg/g for a soil extract with a volume of 50 µl. The assay takes approximately half of an hour and was fully correlated to the Bradford assay. The biosensor had significant advantages than the other methods: it worked in a label-free mode and was fully applicable for practical samples.

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Quantitative Assessment of Periodontal Bacteria Using a Cell-Based Immunoassay with Functionalized Quartz Crystal Microbalance

Chemosensors ◽

10.3390/chemosensors9070159 ◽

2021 ◽

Vol 9 (7) ◽

pp. 159

Author(s):

Satit Rodphukdeekul ◽

Miyuki Tabata ◽

Chindanai Ratanaporncharoen ◽

Yasuo Takeuchi ◽

Pakpum Somboon ◽

...

Keyword(s):

Monoclonal Antibody ◽

Quartz Crystal Microbalance ◽

Quantitative Assessment ◽

Quartz Crystal ◽

Dynamic Range ◽

Gold Surface ◽

Enzyme Linked Immunosorbent Assay ◽

Inflammatory Disorder ◽

Label Free ◽

Periodontal Bacteria

Periodontal disease is an inflammatory disorder that is triggered by bacterial plaque and causes the destruction of the tooth-supporting tissues leading to tooth loss. Several bacteria species, including Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans, are considered to be associated with severe periodontal conditions. In this study, we demonstrated a quartz crystal microbalance (QCM) immunoassay for quantitative assessment of the periodontal bacteria, A. actinomycetemcomitans. An immunosensor was constructed using a self-assembled monolayer of 11-mercaptoundecanoic acid (11-MUA) on the gold surface of a QCM chip. The 11-MUA layer was evaluated using a cyclic voltammetry technique to determine its mass and packing density. Next, a monoclonal antibody was covalently linked to 11-MUA using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide/N-hydroxysuccinimide to act as the biorecognition element. The specificity of the monoclonal antibody was confirmed by an enzyme-linked immunosorbent assay. A calibration curve, for the relationship between the frequency shifts and number of bacteria, was used to calculate the number of A. actinomycetemcomitans bacteria in a test sample. Based on a regression equation, the lower detection limit was 800 cells, with a dynamic range up to 2.32 × 106 cells. Thus, the QCM biosensor in this study provides a sensitive and label-free method for quantitative analysis of periodontal bacteria. The method can be used in various biosensing assays for practical application and routine detection of periodontitis pathogens.

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A Real-Time Method for Improving Stability of Monolithic Quartz Crystal Microbalance Operating under Harsh Environmental Conditions

Sensors ◽

10.3390/s21124166 ◽

2021 ◽

Vol 21 (12) ◽

pp. 4166

Author(s):

Román Fernández ◽

María Calero ◽

Yolanda Jiménez ◽

Antonio Arnau

Keyword(s):

Real Time ◽

Quartz Crystal Microbalance ◽

Quartz Crystal ◽

Limit Of Detection ◽

Quartz Substrate ◽

Low Cost ◽

Discrete Wavelet ◽

Reagent Consumption ◽

Measurement Cell ◽

The Stability

Monolithic quartz crystal microbalance (MQCM) has recently emerged as a very promising technology suitable for biosensing applications. These devices consist of an array of miniaturized QCM sensors integrated within the same quartz substrate capable of detecting multiple target analytes simultaneously. Their relevant benefits include high throughput, low cost per sensor unit, low sample/reagent consumption and fast sensing response. Despite the great potential of MQCM, unwanted environmental factors (e.g., temperature, humidity, vibrations, or pressure) and perturbations intrinsic to the sensor setup (e.g., mechanical stress exerted by the measurement cell or electronic noise of the characterization system) can affect sensor stability, masking the signal of interest and degrading the limit of detection (LoD). Here, we present a method based on the discrete wavelet transform (DWT) to improve the stability of the resonance frequency and dissipation signals in real time. The method takes advantage of the similarity among the noise patterns of the resonators integrated in an MQCM device to mitigate disturbing factors that impact on sensor response. Performance of the method is validated by studying the adsorption of proteins (neutravidin and biotinylated albumin) under external controlled factors (temperature and pressure/flow rate) that simulate unwanted disturbances.

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Sensor Systems for Label-Free Detection of Biomolecular Interactions: Quartz Crystal Microbalance (QCM) and Surface Plasmon Resonance (SPR)

Sensors for Diagnostics and Monitoring ◽

10.1201/9781351250092-8 ◽

2018 ◽

pp. 179-216

Author(s):

Şükran Şeker ◽

M. Taner Vurat ◽

Arin Doğan ◽

A. Eser Elçin ◽

Y. Murat Elçin

Keyword(s):

Surface Plasmon Resonance ◽

Surface Plasmon ◽

Quartz Crystal Microbalance ◽

Plasmon Resonance ◽

Quartz Crystal ◽

Biomolecular Interactions ◽

Sensor Systems ◽

Label Free ◽

Label Free Detection

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Quantification of C60-induced membrane disruption using a quartz crystal microbalance

RSC Advances ◽

10.1039/c7ra13690k ◽

2018 ◽

Vol 8 (18) ◽

pp. 9841-9849 ◽

Cited By ~ 6

Author(s):

Yuxuan Zeng ◽

Qi Wang ◽

Qiu Zhang ◽

Wei Jiang

Keyword(s):

Quartz Crystal Microbalance ◽

Quartz Crystal ◽

Lipid Membrane ◽

Electrostatic Force ◽

Membrane Disruption ◽

Induced Membrane

Fullerene C60 NPs adhere on lipid membrane due to electrostatic force and cause membrane disruption.

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Effects of γ-zein peptides on lipid membrane organization: Quartz crystal microbalance with dissipation and Langmuir monolayer studies

Colloids and Surfaces A Physicochemical and Engineering Aspects ◽

10.1016/j.colsurfa.2019.04.074 ◽

2019 ◽

Vol 574 ◽

pp. 86-93 ◽

Cited By ~ 2

Author(s):

Yuan Zou ◽

Runting Pan ◽

Yingli Liu ◽

Xiao Liu ◽

Xiaowei Chen ◽

...

Keyword(s):

Quartz Crystal Microbalance ◽

Quartz Crystal ◽

Lipid Membrane ◽

Langmuir Monolayer ◽

Membrane Organization

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Rapid and Simultaneous Quantification of 4 Urinary Proteins by Piezoelectric Quartz Crystal Microbalance Immunosensor Array

Clinical Chemistry ◽

10.1373/clinchem.2006.073569 ◽

2006 ◽

Vol 52 (12) ◽

pp. 2273-2280 ◽

Cited By ~ 41

Author(s):

Yang Luo ◽

Ming Chen ◽

Qianjun Wen ◽

Meng Zhao ◽

Bo Zhang ◽

...

Keyword(s):

Quartz Crystal Microbalance ◽

Quartz Crystal ◽

Clinical Chemistry ◽

Label Free ◽

Urinary Proteins ◽

Quartz Crystals ◽

Piezoelectric Quartz ◽

Piezoelectric Immunosensor ◽

Highly Sensitive ◽

Piezoelectric Quartz Crystal

Abstract Background: Urinary proteins are predictive and prognostic markers for diabetes nephropathy. Conventional methods for the quantification of urinary proteins, however, are time-consuming, and most require radioactive labeling. We designed a label-free piezoelectric quartz crystal microbalance (QCM) immunosensor array to simultaneously quantify 4 urinary proteins. Methods: We constructed a 2 × 5 model piezoelectric immunosensor array fabricated with disposable quartz crystals for quantification of microalbumin, α1-microglobulin, β2-microglobulin, and IgG in urine. We made calibration curves after immobilization of antibodies at an optimal concentration and then evaluated the performance characteristics of the immunosensor with a series of tests. In addition, we measured 124 urine samples with both QCM immunosensor array and immunonephelometry to assess the correlation between the 2 methods. Results: With the QCM immunosensor array, we were able to quantify 4 urinary proteins within 15 min. This method had an analytical interval of 0.01–60 mg/L. The intraassay and interassay imprecisions (CVs) were <10%, and the relative recovery rates were 90.3%–109.1%. Nonspecificity of the immunosensor was insignificant (frequency shifts <20 Hz). ROC analyses indicated sensitivities were ≥95.8% and, specificities were ≥76.3%. Bland–Altman difference plots showed the immunosensor array to be highly comparable to immunonephelometry. Conclusions: The QCM system we designed has the advantages of being rapid, label free, and highly sensitive and thus can be a useful supplement to commercial assay methods in clinical chemistry.

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