1. Proteomics for Low Cell Numbers: How to Optimize the Sample Preparation Workflow for Mass Spectrometry Analysis
- Author
-
Kyra van der Pan, Cristina Teodosio, Brigitta A.E. Naber, Paula Díez, Sara Kassem, Anniek L de Jager, Jacques J.M. van Dongen, Alesha Louis, and Inge F de Laat
- Subjects
Proteomics ,Protein digestion ,Computer science ,Sample (material) ,Quantitative proteomics ,Reviews ,Computational biology ,Mass spectrometry ,single-cell proteomics ,01 natural sciences ,Biochemistry ,Workflow ,03 medical and health sciences ,Tandem Mass Spectrometry ,Sample preparation ,paucicellular sample ,030304 developmental biology ,mass spectrometry ,0303 health sciences ,sample preparation ,010401 analytical chemistry ,General Chemistry ,0104 chemical sciences ,Sample collection ,Peptides ,Chromatography, Liquid - Abstract
Nowadays, massive genomics and transcriptomics data can be generated at the single-cell level. However, proteomics in this setting is still a big challenge. Despite the great improvements in sensitivity and performance of mass spectrometry instruments and the better knowledge on sample preparation processing, it is widely acknowledged that multistep proteomics workflows may lead to substantial sample loss, especially when working with paucicellular samples. Still, in clinical fields, frequently limited sample amounts are available for downstream analysis, thereby hampering comprehensive characterization at protein level. To aim at better protein and peptide recoveries, we compare existing and novel approaches in the multistep sample preparation protocols for mass spectrometry studies, from sample collection, cell lysis, protein quantification, and electrophoresis/ staining to protein digestion, peptide recovery, and LC-MS/MS instruments. From this critical evaluation, we conclude that the recent innovations and technologies, together with high quality management of samples, make proteomics on paucicellular samples possible, which will have immediate impact for the proteomics community.
- Published
- 2021