Single-Cell Proteomics: The Critical Role of Nanotechnology

Abstract

In single-cell analysis, biological variability can be attributed to individual cells, their specific state, and the ability to respond to external stimuli, which are determined by protein abundance and their relative alterations. Mass spectrometry (MS)-based proteomics (e.g., SCoPE-MS and SCoPE2) can be used as a non-targeted method to detect molecules across hundreds of individual cells. To achieve high-throughput investigation, novel approaches in Single-Cell Proteomics (SCP) are needed to identify and quantify proteins as accurately as possible. Controlling sample preparation prior to LC-MS analysis is critical, as it influences sensitivity, robustness, and reproducibility. Several nanotechnological approaches have been developed for the removal of cellular debris, salts, and detergents, and to facilitate systematic sample processing at the nano- and microfluidic scale. In addition, nanotechnology has enabled high-throughput proteomics analysis, which have required the improvement of software tools, such as DART-ID or DO-MS, which are also fundamental for addressing key biological questions. Single-cell proteomics has many applications in nanomedicine and biomedical research, including advanced cancer immunotherapies or biomarker characterization, among others; and novel methods allow the quantification of more than a thousand proteins while analyzing hundreds of single cells.

Single-Cell Proteomics with Spatial Attributes: Tools and Techniques

“…84,85 Further, the coupling of nanotechnology with mass spectrometry has made single-cell-type omics more precise. 86,1 The localization of crucial route elements and assemblies that are responsible for triggering changes in the cellular states throughout the initial development or even the advent of diseases like diabetes, cancer, heart failure, or infections could be revealed, most notably, by improvements in spatial proteomics. When the level of sickness is still clinically manageable as well as reversible, single-cell spatial proteomics may even deliver the promise of customized precision medicine for decades to come by offering a rich new stream of mechanistic insights into human cell biology and disease processes.…”

“…There is the use of freeze–heat process foe cell lysis instead of sonication. Scientists were able to identify around 3042 proteins from 1490 single monocytes and macrophages …”

“…Ground-breaking developments in this field are just around the corner and have the capacity to significantly alter the research in the biomedical field in comparison to the single-cell genomics revolution. Single-cell proteomics has immense importance over single-cell transcriptomics in the clinical aspect because disease pathobiology is directly reflected by the translation of faulty protein, associated post-translation modifications and subcellular localization of protein expression. , Further, the coupling of nanotechnology with mass spectrometry has made single-cell-type omics more precise. , …”

“…Therefore, studying them will give us a clearer understanding regarding the pathogenesis of any disease. This will further be helpful in developing therapeutics for the same …”

“…The task becomes more complex because proteins cannot be amplified like nucleic acids. Therefore, there is a need for advanced sampling procedures to reduce the protein loss, and we need highly accurate methods for proteomics analysis …”