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Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification
Published on: November 15, 2017
Novel prefractionation method can be used in proteomic analysis
Haixin Bai1, Fan Yang, Xiurong Yang
1State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Renmin Street 5625, Changchun, Jilin 130022, China.
Abstract:
For the first time, a novel prefractionation method used in proteomic analysis was developed, which is performed by a novel aqueous two-phase system (NATPS) composed of n-butanol, (NH(4))(2)SO(4), and water. It can separate proteomic proteins into multigroups by one-step extraction. The phase-separation conditions of n-butanol solutions were studied in the presence of commonly used inorganic salts. The NATPS was subsequently developed. Using human serum albumin, zein, and gamma-globulin as model proteins, the separation effectiveness of the NATPS for protein was studied under affection factors, i.e., pH, n-butanol volume, protein, or salt concentration. The model and actual protein samples were separated by the NATPS and then directly used for gel electrophoresis without separating the target proteins from phase-forming reagents. It revealed that the NATPS could separate proteomic proteins into multigroups by one-step extraction. The NATPS has the advantages of rapidity, simplicity, low cost, biocompability, and high efficiency. It need not separate target proteins from the phase-forming reagents. The NATPS has great significance in separation and extraction of proteomic proteins, as well as in methodology.
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