Rapid characterization of secreted recombinant proteins by native mass spectrometry. (December 2018)

• Record Type: Journal Article
• Authors: Ben-Nissan, Gili; Vimer, Shay; Warszawski, Shira; Katz, Aliza; Yona, Meital; Unger, Tamar; Peleg, Yoav; Morgenstern, David; Cohen-Dvashi, Hadas; Diskin, Ron; Fleishman, Sarel; Sharon, Michal
• Is Part Of: Communications biology. Volume 1:Number 1(2018)
• Page Start: 1
• Page End: 12
• Publication Date: 2018-12
• Subjects: Systems biology -- Periodicals; 570.113
• DOI: 10.1038/s42003-018-0231-3
• Languages: English
• ISSNs: 2399-3642

Abstract

Abstract Characterization of overexpressed proteins is essential for assessing their quality, and providing input for iterative redesign and optimization. This process is typically carried out following purification procedures that require pronounced cost of time and labor. Therefore, quality assessment of recombinant proteins with no prior purification offers a major advantage. Here, we report a native mass spectrometry method that enables characterization of overproduced proteins directly from culture media. Properties such as solubility, molecular weight, folding, assembly state, overall structure, post-translational modifications and binding to relevant biomolecules are immediately revealed. We show the applicability of the method for in-depth characterization of secreted recombinant proteins from eukaryotic systems such as yeast, insect, and human cells. This method, which can be readily extended to high-throughput analysis, considerably shortens the time gap between protein production and characterization, and is particularly suitable for characterizing engineered and mutated proteins, and optimizing yield and quality of overexpressed proteins. Ben-Nissan et al. present a rapid mass-spectrometry method for characterizing recombinant proteins directly from culture media. They test their method on secreted recombinant proteins from yeast, insect, and human cells, revealing solubility, molecular weight, structure, ligand binding and post-translational modifications.