Imaging label-free biosensor for multiplexed protein detection

Author(s):  
Sabrina Jahns ◽  
Martina Gerken
Keyword(s):  
2007 ◽  
Vol 12 (5) ◽  
pp. 311-317 ◽  
Author(s):  
Vindhya Kunduru ◽  
Shalini Prasad

We demonstrate a technique to detect protein biomarkers contained in vulnerable coronary plaque using a platform-based microelectrode array (MEA). The detection scheme is based on the property of high specificity binding between antibody and antigen similar to most immunoassay techniques. Rapid clinical diagnosis can be achieved by detecting the amount of protein in blood by analyzing the protein's electrical signature. Polystyrene beads which act as transportation agents for the immobile proteins (antigen) are electrically aligned by application of homogenous electric fields. The principle of electrophoresis is used to produce calculated electrokinetic movement among the anti-C-reactive protein (CRP), or in other words antibody funtionalized polystyrene beads. The electrophoretic movement of antibody-functionalized polystyrene beads results in the formation of “Microbridges” between the two electrodes of interest which aid in the amplification of the antigen—antibody binding event. Sensitive electrical equipment is used for capturing the amplified signal from the “Microbridge” which essentially behaves as a conducting path between the two electrodes. The technique circumvents the disadvantages of conventional protein detection methods by being rapid, noninvasive, label-free, repeatable, and inexpensive. The same principle of detection can be applied for any receptor—ligand-based system because the technique is based only on the volume of the analyte of interest. Detection of the inflammatory coronary disease biomarker CRP is achieved at concentration levels spanning over the lower microgram/milliliter to higher order nanogram/milliliter ranges.


2015 ◽  
Vol 357 (1) ◽  
pp. 74-78
Author(s):  
Anjum Qureshi ◽  
Irena Roci ◽  
Yasar Gurbuz ◽  
Javed H. Niazi

2008 ◽  
Vol 129 (1) ◽  
pp. 372-379 ◽  
Author(s):  
Jung A Lee ◽  
Seongpil Hwang ◽  
Juhyoun Kwak ◽  
Se Il Park ◽  
Seung S. Lee ◽  
...  

2014 ◽  
Vol 194 ◽  
pp. 478-483 ◽  
Author(s):  
Hai-Bo Wang ◽  
Hong-Ding Zhang ◽  
Shu-Ping Xu ◽  
Tian Gan ◽  
Ke-Jing Huang ◽  
...  

Sensors ◽  
2021 ◽  
Vol 21 (7) ◽  
pp. 2555
Author(s):  
Nikolay Mukhin ◽  
Georgii Konoplev ◽  
Aleksandr Oseev ◽  
Marc-Peter Schmidt ◽  
Oksana Stepanova ◽  
...  

Diagnostic devices for point-of-care (POC) urine analysis (urinalysis) based on microfluidic technology have been actively developing for several decades as an alternative to laboratory based biochemical assays. Urine proteins (albumin, immunoglobulins, uromodulin, haemoglobin etc.) are important biomarkers of various pathological conditions and should be selectively detected by urinalysis sensors. The challenge is a determination of different oligomeric forms of the same protein, e.g., uromodulin, which have similar bio-chemical affinity but different physical properties. For the selective detection of different types of proteins, we propose to use a shear bulk acoustic resonator sensor with an additional electrode on the upper part of the bioliquid-filled channel for protein electric field manipulation. It causes modulation of the protein concentration over time in the near-surface region of the acoustic sensor, that allows to distinguish proteins based on their differences in diffusion coefficients (or sizes) and zeta-potentials. Moreover, in order to improve the sensitivity to density, we propose to use structured sensor interface. A numerical study of this approach for the detection of proteins was carried out using the example of albumin, immunoglobulin, and oligomeric forms of uromodulin in model urine solutions. In this contribution we prove the proposed concept with numerical studies for the detection of albumin, immunoglobulin, and oligomeric forms of uromodulin in urine models.


2016 ◽  
Vol 8 (21) ◽  
pp. 4177-4180 ◽  
Author(s):  
Bin Wu ◽  
Nandi Chen ◽  
Qing Wang ◽  
Xiaohai Yang ◽  
Kemin Wang ◽  
...  

We developed a simple label-free aptamer-based method for C-reactive protein (CRP) detection.


2012 ◽  
Vol 1414 ◽  
Author(s):  
Rahim Esfandyarpour ◽  
Hesaam Esfandyarpour ◽  
Mehdi Javanmard ◽  
James S. Harris ◽  
Ronald W. Davis

Abstract:Here we present the development of an array of electrical nano-biosensors in a microfluidic channel, called Nanoneedle biosensors. Then we present the proof of concept study for protein detection. A Nanoneedle biosensor is a real-time, label-free, direct electrical detection platform, which is capable of high sensitivity detection, measuring the change in ionic current and impedance modulation, due to the presence or reaction of biomolecules such as proteins or nucleic acids. We show that the sensors which have been fabricated and characterized for the protein detection. We have functionalized Nanoneedle biosensors with receptors specific to a target protein using physical adsorption for immobilization. We have used biotinylated bovine serum albumin as the receptor and sterptavidin as the target analyte. The detection of streptavidin binding to the receptor protein is also presented.


2020 ◽  
Author(s):  
J. Peter Rickgauer ◽  
Heejun Choi ◽  
Jennifer Lippincott-Schwartz ◽  
Winfried Denk

AbstractA general method to map molecular interactions and conformational states in structurally intact cells would find wide application in biochemistry and cell biology. We used a library of images— calculated on the basis of known structural data—as search templates to detect targets as small as the “head” domain (350 kDa) of the ribosome’s small subunit in single-tilt electron cryo-micrographs by cellular high resolution template matching (cHRTM). Atomically precise position and orientation estimates reveal the conformation of individual ribosomes and enable the detection of specifically bound ligands down to 24 kDa. We show that highly head-swivelled states are likely to play a role in mRNA translocation in living cells. cHRTM outperforms cryo-electron tomography three-fold in sensitivity and completely avoids the vicissitudes of exogenous labelling.


F1000Research ◽  
2014 ◽  
Vol 2 ◽  
pp. 272 ◽  
Author(s):  
Jakob Vowinckel ◽  
Floriana Capuano ◽  
Kate Campbell ◽  
Michael J. Deery ◽  
Kathryn S. Lilley ◽  
...  

The combination of qualitative analysis with label-free quantification has greatly facilitated the throughput and flexibility of novel proteomic techniques. However, such methods rely heavily on robust and reproducible sample preparation procedures. Here, we benchmark a selection of in gel, on filter, and in solution digestion workflows for their application in label-free proteomics. Each procedure was associated with differing advantages and disadvantages. The in gel methods interrogated were cost effective, but were limited in throughput and digest efficiency. Filter-aided sample preparations facilitated reasonable processing times and yielded a balanced representation of membrane proteins, but led to a high signal variation in quantification experiments. Two in solution digest protocols, however, gave optimal performance for label-free proteomics. A protocol based on the detergent RapiGest led to the highest number of detected proteins at second-best signal stability, while a protocol based on acetonitrile-digestion, RapidACN, scored best in throughput and signal stability but came second in protein identification. In addition, we compared label-free data dependent (DDA) and data independent (SWATH) acquisition on a TripleTOF 5600 instrument. While largely similar in protein detection, SWATH outperformed DDA in quantification, reducing signal variation and markedly increasing the number of precisely quantified peptides.


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