公开(公告)号：US11365214B2

公开(公告)日：2022-06-21

申请号：US16462464

申请日：2017-07-12

申请人： DALIAN INSTITUTE OF CHEMICAL PHYSICS, CHINESE ACADEMY OF SCIENCES

发明人： Lihua Zhang ,  Huiming Yuan ,  Zhigang Sui ,  Kaiguang Yang ,  Yukui Zhang

IPC分类号： C07K1/14 ,  C12Q1/37 ,  G01N1/40 ,  G01N30/14 ,  G01N30/72 ,  G01N30/06

摘要： The invention relates to a method for the treatment of body fluid proteins, by which proteins from body fluids such as blood or urine are extracted by adding a certain proportion of high molecular polymer solution under low temperature condition followed by denaturation and reduction by adding a certain concentration of surfactant and tris(2-carboxyethyl) phosphine (TCEP) under a high temperature condition. Subsequently, the iodoacetic acid brushes grafted on silica microspheres called as solid-phase alkylation reagents are added into protein solution, which can react rapidly with the protein sulfhydryl group. After centrifugation, the microspheres are obtained and repeatedly washed with methanol and buffer to remove interferences such as sugars, salts, surfactants, lipids to obtain high-purity proteins, and finally protease is added to digest proteins into peptides. After centrifugation, the peptide products are obtained, and directly analyzed by liquid chromatography-mass spectrometry (LC-MS) system. Compared with the traditional protein pretreatment method, the method has many advantages such as good anti-interference capability, easy operation and short pretreatment time.