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Proteins and Proteomics
Published in Firdos Alam Khan, Biotechnology Fundamentals, 2020
A protein microarray, sometimes referred to as a protein binding microarray, provides a multiplex approach to identify protein–protein interactions, to identify the substrates of protein kinases, to identify TF protein-activation, or to identify the targets of biologically active small molecules. The array is a piece of glass on which different molecules of protein or specific DNA binding sequences (as capture probes for the proteins) have been affixed at separate locations in an ordered manner thus forming a microscopic array. The most common protein microarray is the antibody microarray, where antibodies are spotted onto the protein chip and are used as capture molecules to detect proteins from cell lysate solutions. Protein microarrays (also biochip, protein chip) are measurement devices used in biomedical applications to determine the presence and/or amount (referred to as relative quantitation) of proteins in biological samples, for example, blood. They have the potential to be an important tool for proteomics research. Usually different capture agents, most frequently monoclonal antibodies, are deposited on a chip surface (glass or silicon) in a miniature array. This format is often referred to as a microarray (a more general term for chip-based biological measurement devices). There are several types of protein chips, the most common being glass slide chips and nano-well arrays (Figure 3.18).
Proteins and proteomics
Published in Firdos Alam Khan, Biotechnology Fundamentals, 2018
A protein microarray, sometimes referred to as a protein-binding microarray, provides a multiplex approach to identify protein—protein interactions, to identify the substrates of protein kinases, to identify TF protein-activation, or to identify the targets of biologically active small molecules. The array is a piece of glass on which different molecules of protein or specific DNA-binding sequences (as capture probes for the proteins) have been affixed at separate locations in an ordered manner, thus forming a microscopic array. The most common protein microarray is the antibody microarray, where antibodies are spotted onto the protein chip and are used as capture molecules to detect proteins from cell lysate solutions. Protein microarrays (also biochip, proteinchip) are measurement devices used in biomedical applications to determine the presence and/or amount (referred to as relative quantitation) of proteins in biological samples, for example, blood. They have the potential to be an important tool for proteomics research. Usually different capture agents, most frequently monoclonal antibodies, are deposited on a chip surface (glass or silicon) in a miniature array. This format is often referred to as a microarray (a more general term for chip-based biological measurement devices). There are several types of protein chips, the most common being glass slide chips and nano-well arrays (Figure 3.18).
Biomems
Published in Simona Badilescu, Muthukumaran Packirisamy, BioMEMS, 2016
Simona Badilescu, Muthukumaran Packirisamy
In the postgenomic era, proteomics has enormous potential in biology and medicine. In spite of the technical problems, protein microarray technology has emerged as a promising approach for profiling the protein expression in diagnostic, prognostic, and disease progression monitoring. Some of the applications of protein microarray include identification of protein-protein interactions, protein-phospholipid interactions, small-molecule targets, and substrates of protein kinases. In order to provide insights into the mechanisms of biological processes and to get detailed information about a complex cellular system, the identification and analysis of each of its components and the determination of their biochemical activities are required. The most common protein microarray is the antibody microarray, as shown in Figure 9.23, where antibodies are spotted onto the chip and used as capture molecules to detect proteins from cells.
Emerging applications of microfluidic techniques for in vitro toxicity studies of atmospheric particulate matter
Published in Aerosol Science and Technology, 2021
Fobang Liu, Nga Lee Ng, Hang Lu
Recently, several microfluidic devices have been developed and applied for high-throughput and high-content analysis of multiple cellular endpoints upon PM exposure. Zheng et al. (2017) designed and fabricated a microfluidic system consisting of concentration gradient generators, cell culture chambers, and an immune-detection chip. The microfluidic system enables the processes of PM dilution, cell culture, cell stimulation, and immunoassay to be done on-chip. Figure 3 shows a schematic of the microfluidic system, which is comprised of twelve uniform cell culture chips and an immunodetection chip. Each cell culture chip contains a concentration gradient generator and parallel cell culture chambers with a perfusion module. The immunodetection chip is used for protein microarray assays. A typical antibody “Sandwich” immunoassay is introduced to this system with all steps operated on the device. The protein analysis results of the on‐chip immunoassay show a good correlation with results from traditional ELISA. Importantly, the system requires a minimal amount of PM mass and could detect a large number of proteins. The authors demonstrated that the required PM mass for their system is around three orders of magnitude smaller than that for traditional ELISA (1.8 µg vs. 1.2 mg). Zhang et al. (2017) fabricated a similar microfluidic system that integrates a cell culture chip and protein microarray chip to investigate the mechanisms of PM-mediated cytotoxicity in human skin cells. In the study of Cui et al. (2015), they also developed a microfluidic system consisting of a concentration gradient generator and cell culture chambers, which is capable of conducting on-chip immunofluorescent analysis. Besides that, the cell culture chamber module is comprised of two layers, in which an upper layer is for introducing human alveolar macrophages and a lower layer is for introducing human bronchial cells (16HBE). This design is to enable studying the effect of PM-treated 16HBE cells on alveolar macrophage migration.