TY - JOUR
T1 - Rapid characterization of secreted recombinant proteins by native mass spectrometry
AU - Ben-Nissan, Gili
AU - Vimer, Shay
AU - Warszawski, Shira
AU - Katz, Aliza
AU - Yona, Meital
AU - Unger, Tamar
AU - Peleg, Yoav
AU - Morgenstern, David
AU - Cohen-Dvashi, Hadas
AU - Diskin, Ron
AU - Fleishman, Sarel J
AU - Sharon, Michal
N1 - We thank Ely Morag and Ed Bayer (Weizmann Institute of Science) for providing us with the pPICK9 plasmid for CBM3a protein expression. M.S. is also grateful for the support of a Starting Grant from the European Research Council (ERC) (Horizon 2020)/ERC Grant Agreement No. 636752 and for an Israel Science Foundation (ISF) grant 300/17. M.S. is an incumbent of the Aharon and Ephraim Katzir Memorial Professorial Chair. These authors contributed equally: Gili Ben-Nissan, Shay Vimer. G.B.N., S.V. and M.S. designed the experiments. GBN and SV performed the experiments. G.B.N., S.V. and M.S. and analyzed the data with assistance from S.W. A.K., M.Y., T.U., Y.P., D.M., H.C.D., R.D. and S.J.F. G.B.N., S.V. and M.S. wrote and edited the paper.
PY - 2018/12/3
Y1 - 2018/12/3
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85071169329&partnerID=8YFLogxK
U2 - https://doi.org/10.1038/s42003-018-0231-3
DO - https://doi.org/10.1038/s42003-018-0231-3
M3 - مقالة
C2 - 30534605
SN - 2399-3642
VL - 1
JO - Communications Biology
JF - Communications Biology
IS - 1
M1 - 213
ER -