The DOI matches its Crossref record; 143 of the 143 references it cites were found in a registry or an index.
Proteomic Challenges: Sample Preparation Techniques for Microgram-Quantity Protein Analysis from Biological Samples
Open accessAbstract
Proteins regulate many cellular functions and analyzing the presence and abundance of proteins in biological samples are central focuses in proteomics. The discovery and validation of biomarkers, pathways, and drug targets for various diseases can be accomplished using mass spectrometry-based proteomics. However, with mass-limited samples like tumor biopsies, it can be challenging to obtain sufficient amounts of proteins to generate high-quality mass spectrometric data. Techniques developed for macroscale quantities recover sufficient amounts of protein from milligram quantities of starting material, but sample losses become crippling with these techniques when only microgram amounts of material are available. To combat this challenge, proteomicists have developed micro-scale techniques that are compatible with decreased sample size (100 μg or lower) and still enable excellent proteome coverage. Extraction, contaminant removal, protein quantitation, and sample handling techniques for the microgram protein range are reviewed here, with an emphasis on liquid chromatography and bottom-up mass spectrometry-compatible techniques. Also, a range of biological specimens, including mammalian tissues and model cell culture systems, are discussed.
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Cites
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Distinct mechanisms of microRNA sorting into cancer cell-derived extracellular vesicle subtypes
Temoche-Diaz, M. M., Shurtleff, M. J., Nottingham, R. M., Yao, J., Fadadu, R. P., Lambowitz, A. M., & Schekman, R. (2019) · eLife · doi:10.7554/elife.47544 · Cited by 227
Edwards, J. E., Forster, R. J., Callaghan, T. M., Dollhofer, V., Dagar, S. S., Cheng, Y., Chang, J., Kittelmann, S., Fliegerova, K., Puniya, A. K., et al. (2017) · Frontiers in Microbiology · doi:10.3389/fmicb.2017.01657 · Cited by 130
Cell-type-specific metabolic labeling, detection and identification of nascent proteomes in vivo
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Kwiatkowski, M., Wurlitzer, M., Krutilin, A., Kiani, P., Nimer, R., Omidi, M., Mannaa, A., Bussmann, T., Bartkowiak, K., Kruber, S., et al. (2016) · Journal of Proteomics · doi:10.1016/j.jprot.2015.12.029 · Cited by 43
Metadata
- Authors
- Feist, P., & Hummon, A.
- Venue
- International Journal of Molecular Sciences
- Publisher
- MDPI AG
- Published
- 2015-02-05
- Volume
- 16
- Issue
- 2
- Pages
- 3537-3563
- ISSN
- 1422-0067 (ISSN-L)
- OpenAlex
- W2066387545
- Cited by
- 288
- References
- 143
- FWCI
- 11.89
- Type
- journal article
- Language
- en
- Open access
- gold
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