Protein and Proteome Measurements with Microfluidic Chips

Abstract

Major scientific discoveries in the last two decades have been enabled by the development of high-throughput sequencing and mass spectrometry (MS) technologies. Omics analysis, at the genome, transcriptome, proteome, or metabolome levels, has paved the way for the elucidation of many molecular mechanisms that underlie disease, biomarker and drug-target discovery, and diagnostics and precision medicine applications. With the realization that the biological interpretation of results generated from biological tissues or cell-derived samples is highly impacted by cell heterogeneity, many efforts have been redirected toward the omics characterization of single cells. As miniaturized devices represent ideal platforms for the analysis of small sample amounts, omics analysis has found a flourishing ground in the microfluidics instrumentation development arena. This Review aims at capturing the latest protein and proteome analysis technologies that have been developed and implemented on microfluidic platforms, with a focus on developments that have been documented in the past two years. Emphasis was placed on challenging aspects that relate to sensitivity and handling sample complexity, as well as on applications that address demanding biological and biomedical problems.

Oxygen control: the often overlooked but essential piece to create betterin vitrosystems

“…Multichannel (cyclical) single molecule FISH (fluorescence in situ hybridization) techniques could be used for transcriptome level measurements while retaining the spatial resolution, which is key to correlate gene expression profile and local oxygen abundance. 157 Other approaches that have been developed for off-chip analyses include the determination of the cytokine secretion profile (secretomics) 158 and their metabolic activity using collected medium (metabolomics), 159 or their protein expression profile (proteomics), 160 as well as gene expression analysis or single-cell transcriptomic studies, notably single cell RNA sequencing, after recovery of the cellular materials. 161…”

Microfluidics for Peptidomics, Proteomics, and Cell Analysis

“…Sonker et al [ 110 ] reported the possibility of using a specific separation system for integrated immunoaffinity extraction to study human serum matrix biomarkers in preterm infants. They used a reactive polymer to immobilize the antibody as a whole and selectively extracted targeted preterm infant markers.…”

“…It is expected that the specific cells in the biopsy sample will be treated individually. Further analysis should focus on system expansion, which obviously requires thorough optimization [ 110 ].…”

Microfluidic Device with an Integrated Freeze-Dried Cell-Free Protein Synthesis System for Small-Volume Biosensing

“…This advantage is useful to reduce invasiveness in liquid biopsy using blood, urine, saliva, and tears. Therefore, microfluidic devices have been applied as point-of-care (PoC) diagnostic devices [ 1 , 2 , 3 , 4 ]. Recently, an attractive PoC device has been developed [ 5 ] (i.e., a microfluidic device equipped with freeze-dried diagnostic reagents).…”