scholarly journals Protein detection in blood with single-molecule imaging

2021 ◽  
Vol 7 (33) ◽  
pp. eabg6522
Author(s):  
Chih-Ping Mao ◽  
Shih-Chin Wang ◽  
Yu-Pin Su ◽  
Ssu-Hsueh Tseng ◽  
Liangmei He ◽  
...  
Keyword(s):  
Single Molecule ◽  
Imaging Technique ◽  
Protein Detection ◽  
Single Molecule Imaging ◽  
Blood Samples ◽  
Intracellular Proteins ◽  
Protein Molecules ◽  
Human Blood Samples ◽  
Disease Profiling ◽  
Diagnosis Of Diseases

The ability to characterize individual biomarker protein molecules in patient blood samples could enable diagnosis of diseases at an earlier stage, when treatment is typically more effective. Single-molecule imaging offers a promising approach to accomplish this goal. However, thus far, single-molecule imaging methods have not been translated into the clinical setting. The detection limit of these methods has been confined to the picomolar (10−12 M) range, several orders of magnitude higher than the circulating concentrations of biomarker proteins present in many diseases. Here, we describe single-molecule augmented capture (SMAC), a single-molecule imaging technique to quantify and characterize individual protein molecules of interest down to the subfemtomolar (<10−15 M) range. We demonstrate SMAC in a variety of applications with human blood samples, including the analysis of disease-associated secreted proteins, membrane proteins, and rare intracellular proteins. SMAC opens the door to the application of single-molecule imaging in noninvasive disease profiling.

scholarly journals Protein detection in blood with single-molecule imaging

2021 ◽  
Author(s):  
Chic-Ping Mao ◽  
Shih-Chin Wang ◽  
Yu-Pin Su ◽  
Ssu-Hsueh Tseng ◽  
Liangmei He ◽  
...  
Keyword(s):  
Single Molecule ◽  
Early Stage ◽  
Protein Detection ◽  
Tp53 Gene ◽  
Invasive Disease ◽  
Structural Features ◽  
Single Molecule Imaging ◽  
Blood Samples ◽  
Mutant Proteins ◽  
Protein Molecules

The ability to identify and characterize individual biomarker protein molecules in patient blood samples could enable diagnosis of diseases at an earlier stage, when treatment is typically more effective. Single-molecule imaging offers a promising approach to accomplish this goal. However, thus far single-molecule imaging methods have only been used to monitor protein molecules in solutions or cell lysates, and have not been translated into the clinical arena. Furthermore, the detection limit of these methods has been confined to the picomolar (10-12 M) range. In many diseases, the circulating concentrations of biomarker proteins fall several orders of magnitude below this range. Here we describe Single-Molecule Augmented Capture (SMAC), a single-molecule imaging technique to visualize, quantify, and characterize individual protein molecules of interest down to the subfemtomolar (<10-15 M) range, even in complex biologic fluids. We demonstrate SMAC in a wide variety of applications with human blood samples, including the analysis of disease-associated secreted proteins, membrane proteins, and rare intracellular proteins. Using ovarian cancer as a model, a lethal malignancy in which high-grade disease is driven almost universally by alterations in the TP53 gene and frequently only diagnosed at a late, incurable stage, we found that mutant pattern p53 proteins are released into the blood in patients at an early stage in disease progression. SMAC opens the door to the application of single-molecule imaging in non-invasive disease profiling and allows for the analysis of circulating mutant proteins as a new class of highly specific disease biomarkers. The SMAC platform can be adapted to multiplex or high-throughput formats to characterize heterogeneous biochemical and structural features of circulating proteins-of-interest.

scholarly journals Protein Detection in Blood with Single-Molecule Imaging

2019 ◽  
Vol 116 (3) ◽  
pp. 465a
Author(s):  
Shih-Chin Wang ◽  
Chih-Ping Mao ◽  
Yu-Pin Su ◽  
T.C. Wu ◽  
Chien-Fu Hung ◽  
...  
Keyword(s):  
Single Molecule ◽  
Protein Detection ◽  
Single Molecule Imaging

scholarly journals Application of Solid-State Nanopore in Protein Detection

2020 ◽  
Vol 21 (8) ◽  
pp. 2808 ◽  
Author(s):  
Yuhan Luo ◽  
Linlin Wu ◽  
Jing Tu ◽  
Zuhong Lu
Keyword(s):  
Solid State ◽  
Chemical Modification ◽  
Single Molecule ◽  
Protein Detection ◽  
High Sensitivity ◽  
Protein Sequencing ◽  
Its Sequence ◽  
Sequence Structure ◽  
Single Molecule Level ◽  
Protein Molecules

A protein is a kind of major biomacromolecule of life. Its sequence, structure, and content in organisms contains quite important information for normal or pathological physiological process. However, research of proteomics is facing certain obstacles. Only a few technologies are available for protein analysis, and their application is limited by chemical modification or the need for a large amount of sample. Solid-state nanopore overcomes some shortcomings of the existing technology, and has the ability to detect proteins at a single-molecule level, with its high sensitivity and robustness of device. Many works on detection of protein molecules and discriminating structure have been carried out in recent years. Single-molecule protein sequencing techniques based on solid-state nanopore are also been proposed and developed. Here, we categorize and describe these efforts and progress, as well as discuss their advantages and drawbacks.

Single Molecule Imaging of Protein Molecules in Nanopores

2010 ◽  
Vol 82 (2) ◽  
pp. 478-482 ◽  
Author(s):  
Changbei Ma ◽  
Edward S. Yeung
Keyword(s):  
Single Molecule ◽  
Single Molecule Imaging ◽  
Protein Molecules

Label-free, mass-sensitive single-molecule imaging using interferometric scattering microscopy

2020 ◽  
Author(s):  
Nikolas Hundt
Keyword(s):  
Single Molecule ◽  
Cellular Systems ◽  
Single Molecule Imaging ◽  
Label Free ◽  
Measurement Sensitivity ◽  
Mass Distributions ◽  
Quantitative Measurements ◽  
Contrast Mechanism ◽  
Cellular Components ◽  
Scattering Signal

Abstract Single-molecule imaging has mostly been restricted to the use of fluorescence labelling as a contrast mechanism due to its superior ability to visualise molecules of interest on top of an overwhelming background of other molecules. Recently, interferometric scattering (iSCAT) microscopy has demonstrated the detection and imaging of single biomolecules based on light scattering without the need for fluorescent labels. Significant improvements in measurement sensitivity combined with a dependence of scattering signal on object size have led to the development of mass photometry, a technique that measures the mass of individual molecules and thereby determines mass distributions of biomolecule samples in solution. The experimental simplicity of mass photometry makes it a powerful tool to analyse biomolecular equilibria quantitatively with low sample consumption within minutes. When used for label-free imaging of reconstituted or cellular systems, the strict size-dependence of the iSCAT signal enables quantitative measurements of processes at size scales reaching from single-molecule observations during complex assembly up to mesoscopic dynamics of cellular components and extracellular protrusions. In this review, I would like to introduce the principles of this emerging imaging technology and discuss examples that show how mass-sensitive iSCAT can be used as a strong complement to other routine techniques in biochemistry.

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