Nanoproteomics comes of age

Abstract

Nanoproteomics, which is defined as quantitative proteome profiling of small populations of cells (<5000 cells), can reveal critical information related to rare cell populations, hard-to-obtain clinical specimens, and the cellular heterogeneity of pathological tissues. Areas covered: We present a brief review of the recent technological advances in nanoproteomics. These advances include new technologies or approaches covering major areas of proteomics workflow ranging from sample isolation, sample processing, high-resolution separations, to MS instrumentation. Expert commentary: We comment on the current state of nanoproteomics and discuss perspectives on both future technological directions and potential enabling applications.

Keywords: Nanoproteomics; nanoPOTS; single cell proteomics.

Figure 1.. Typical workflow for bottom-up nanoproteomics.

Briefly, specified cell population is isolated from tissues or cultured cells. Proteins are extracted and digested by proteolytic enzymes (e.g., trypsin) into peptides. The digested samples are analyzed by nanoflow LC or capillary electrophoresis (CE) coupled with MS through electrospray ionization. Peptide sequences are identified from MS/MS spectra through database searching, and MS intensity information is typically utilized for protein quantification.