scholarly journals A207 Development of membrane protein detection methods on bone cells using AFM

2013 ◽  
Vol 2013.24 (0) ◽  
pp. 61-62
Author(s):  
Takato TAMAKI ◽  
Sung-Woong HAN ◽  
Taiji ADACHI
Keyword(s):  
Membrane Protein ◽  
Bone Cells ◽  
Protein Detection ◽  
Detection Methods

scholarly journals Electrokinetic Formation of “Microbridges” for Protein Biomarkers as Sensors

2007 ◽  
Vol 12 (5) ◽  
pp. 311-317 ◽  
Author(s):  
Vindhya Kunduru ◽  
Shalini Prasad
Keyword(s):  
Electric Fields ◽  
Microelectrode Array ◽  
Protein Detection ◽  
High Specificity ◽  
Detection Methods ◽  
Label Free ◽  
Protein Biomarkers ◽  
Detection Scheme ◽  
Polystyrene Beads ◽  
Conducting Path

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.

scholarly journals Printed Electrodes in Microfluidic Arrays for Cancer Biomarker Protein Detection

Biosensors ◽  
2020 ◽  
Vol 10 (9) ◽  
pp. 115
Author(s):  
Lasangi Dhanapala ◽  
Colleen E. Krause ◽  
Abby L. Jones ◽  
James F. Rusling
Keyword(s):  
Cancer Progression ◽  
Simultaneous Detection ◽  
Protein Detection ◽  
Medical Diagnostics ◽  
Cancer Diagnostics ◽  
Detection Methods ◽  
Enzyme Linked Immunosorbent Assay ◽  
Grand Challenge ◽  
Patient Records ◽  
Personalized Care

Medical diagnostics is trending towards a more personalized future approach in which multiple tests can be digitized into patient records. In cancer diagnostics, patients can be tested for individual protein and genomic biomarkers that detect cancers at very early stages and also be used to monitor cancer progression or remission during therapy. These data can then be incorporated into patient records that could be easily accessed on a cell phone by a health care professional or the patients themselves on demand. Data on protein biomarkers have a large potential to be measured in point-of-care devices, particularly diagnostic panels that could provide a continually updated, personalized record of a disease like cancer. Electrochemical immunoassays have been popular among protein detection methods due to their inherent high sensitivity and ease of coupling with screen-printed and inkjet-printed electrodes. Integrated chips featuring these kinds of electrodes can be built at low cost and designed for ease of automation. Enzyme-linked immunosorbent assay (ELISA) features are adopted in most of these ultrasensitive detection systems, with microfluidics allowing easy manipulation and good fluid dynamics to deliver reagents and detect the desired proteins. Several of these ultrasensitive systems have detected biomarker panels ranging from four to eight proteins, which in many cases when a specific cancer is suspected may be sufficient. However, a grand challenge lies in engineering microfluidic-printed electrode devices for the simultaneous detection of larger protein panels (e.g., 50–100) that could be used to test for many types of cancers, as well as other diseases for truly personalized care.

scholarly journals Evaluation and Validation of Plasma Proteins Using Two Different Protein Detection Methods for Early Detection of Colorectal Cancer

Cancers ◽  
2019 ◽  
Vol 11 (10) ◽  
pp. 1426 ◽  
Author(s):  
Megha Bhardwaj ◽  
Anton Gies ◽  
Korbinian Weigl ◽  
Kaja Tikk ◽  
Axel Benner ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Early Detection ◽  
Plasma Proteins ◽  
Multiple Reaction Monitoring ◽  
Protein Detection ◽  
Detection Methods ◽  
Screening Colonoscopy ◽  
Receptor Protein ◽  
Advanced Adenoma ◽  
Independent Validation

Objective: Plasma protein biomarkers could be an efficient alternative for population-based screening for early detection of colorectal cancer (CRC). The objective of this study was to evaluate and validate plasma proteins individually and as a signature for early detection of CRC. Methods: In a three-stage design, proteins were measured firstly by liquid chromatography/multiple reaction monitoring-mass spectrometry (LC/MRM-MS) and later by proximity extension assay (PEA) in a discovery set consisting of 96 newly diagnosed CRC cases and 94 controls free of neoplasms at screening colonoscopy. Two algorithms (one for each measurement method) were derived by Lasso regression and .632+ bootstrap based on 11 proteins that were included in both the LC/MRM-MS and PEA measurements. Additionally, another algorithm was constructed from the same eleven biomarkers plus amphireglin, the most promising protein marker in the PEA measurements that had not been available from the LC/MRM-MS measurements. Lastly the three prediction signatures were validated with PEA in independent samples of participants of screening colonoscopy (CRC (n = 56), advanced adenoma (n = 101), and participants free of neoplasm (n = 102)). Results: The same four proteins were included in all three prediction signatures; mannan binding lectin serine protease 1, osteopontin, serum paraoxonase lactonase 3 and transferrin receptor protein 1, and the third prediction signature additionally included amphiregulin. In the independent validation set from a true screening setting, the five-marker blood-based signature including AREG presented areas under the curves of 0.82 (95% CI, 0.74–0.89), 0.86 (95% CI, 0.77–0.92) and 0.76 (95% CI, 0.64–0.86) for all, early and late stages CRC, respectively. Conclusion: Two different measurement methods consistently identified four protein markers and an algorithm additionally including amphiregulin, a marker measured by PEA only, showed promising performance for detecting early stage CRC in an independent validation in a true screening setting. These proteins may be potential candidates for blood-based tests for early detection of CRC.

scholarly journals Improved protease digestion conditions for membrane protein detection

2015 ◽  
Vol 36 (15) ◽  
pp. 1690-1698 ◽  
Author(s):  
Lie Min ◽  
Leila H. Choe ◽  
Kelvin H. Lee
Keyword(s):  
Membrane Protein ◽  
Protein Detection ◽  
Protease Digestion

Protein Detection Methods in Proteomics Research

2005 ◽  
Vol 25 (1-2) ◽  
pp. 19-32 ◽  
Author(s):  
Reiner Westermeier ◽  
Rita Marouga
Keyword(s):  
Fluorescent Dyes ◽  
Organic Dyes ◽  
Protein Detection ◽  
Detection Methods ◽  
Mass Determination ◽  
Chemical Methods ◽  
Physical Methods ◽  
Proteomics Research ◽  
Dimensional Electrophoresis

In proteomics research chemical as well as physical methods are used to detect proteins subsequently to their separation. Physical methods are mostly applied after chromatography. They are either based on spectroscopy like light absorption at certain wavelengths or mass determination of peptides and their fragments with mass spectrometry. Chemical methods are used after two-dimensional electrophoresis and employ staining with organic dyes, metal chelates, fluorescent dyes, complexing with silver, or pre-labeling with fluorophores. In some cases autoradiography is still used. Since all of these techniques are very different in terms of sensitivity, their usefulness for quantitative determinations varies significantly. This review will describe the various protein detection methods applied to electrophoresis gels.

scholarly journals AFM-based technologies as the way towards the reverse Avogadro number

2015 ◽  
Vol 61 (2) ◽  
pp. 239-253 ◽  
Author(s):  
T.O. Pleshakova ◽  
I.D. Shumov ◽  
Yu.D. Ivanov ◽  
K.A. Malsagova ◽  
A.L. Kaysheva ◽  
...  
Keyword(s):  
Detection System ◽  
Biological Fluids ◽  
Chemical Properties ◽  
Protein Detection ◽  
Detection Methods ◽  
Practical Implementation ◽  
Advantages And Disadvantages ◽  
Avogadro Number ◽  
Modern Development ◽  
Concentration Detection Limit

Achievement of the concentration detection limit for proteins at the level of the reverse Avogadro number determines the modern development of proteomics. In this review, the possibility of approximating the reverse Avogadro number by using nanotechnological methods (AFM-based fishing with mechanical and electrical stimulation, nanowire detectors, and other methods) are discussed. The ability of AFM to detect, count, visualize and characterize physico-chemical properties of proteins at concentrations up to 10-17-10-18 M is demonstrated. The combination of AFM-fishing with mass-spectrometry allows the identification of proteins not only in pure solutions, but also in multi-component biological fluids (serum). The possibilities to improve the biospecific fishing efficiency by use of SOMAmers in both AFM and nanowire systems are discussed. The paper also provides criteria for evaluation of the sensitivity of fishing-based detection systems. The fishing efficiency depending on the detection system parameters is estimated. The practical implementation of protein fishing depending on the ratio of the sample solution volume and the surface of the detection system is discussed. The advantages and disadvantages of today's promising nanotechnological protein detection methods implemented on the basis of these schemes.

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