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学科主题物理化学
Single Chain Variable Fragment Displaying M13 Phage Library Functionalized Magnetic Microsphere-Based Protein Equalizer for Human Serum Protein Analysis
Zhu, Guijie1; Zhao, Peng1; Deng, Nan1,2; Tao, Dingyin1; Sun, Liangliang1; Liang, Zhen1; Zhang, Lihua1; Zhang, Yukui1; Zhang LH(张丽华)
刊名ANALYTICAL CHEMISTRY
2012-09-18
DOI10.1021/ac3017746
84期:18页:7633-7637
收录类别SCI
文章类型Article
部门归属18
项目归属1810
产权排名1,1
类目[WOS]Chemistry, Analytical
研究领域[WOS]Chemistry
关键词[WOS]PEPTIDE LIGAND LIBRARIES ; LOW-ABUNDANCE PROTEOME ; IN-DEPTH EXPLORATION ; CHROMATOGRAPHY M-LAC ; RED-BLOOD-CELLS ; LIQUID-CHROMATOGRAPHY ; PLASMA-PROTEINS ; TOF-MS ; DEPLETION ; ENRICHMENT
英文摘要Single chain variable fragment (scFv) displaying the M13 phage library was covalently immobilized on magnetic microspheres and used as a protein equalizer for the treatment of human serum. First, scFv displaying M13 phage library functionalized magnetic microspheres (scFv@M13@MM) was incubated with a human serum sample. Second, captured proteins on scFv@M13@MM were eluted with 2 M NaCl, SO mM glycine-hydrochloric acid (Gly-HCl), and 20% (v/v) acetonitrile with 0.5% (v/v) trifluoroacetic acid in sequence. Finally, the tightly bonded proteins were released by the treatment with thrombin. The eluates were first analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with silver staining. Results indicated that the difference of protein concentration was reduced obviously in NaCl and Gly-HCl fractions compared with untreated human serum sample. The eluates were also digested with trypsin, followed by online 2D-strong cation exchange (SCX)-RPLC-ESI-MS/MS analysis. Results demonstrated that the number of proteins identified from an scFv@M13@MM treated human serum sample was improved 100% compared with that from the untreated sample. In addition, the spectral count of 10 high abundance proteins (serum albumin, serotransferrin, alpha-2-macroglobulin, alpha-1-antitrypsin, apolipoprotein B-100, Ig gamma-2 chain C region, haptoglobin, hemopodn, alpha-1-acid glycoprotein 1, and alpha-2-HS-glycoprotein) decreased evidently after scFv@M13@MM treatment. All these results demonstrate that scFv@M13@MM could efficiently remove high-abundance proteins, reduce the protein concentration difference of human serum, and result in more protein identification.
语种英语
WOS记录号WOS:000308829700009
引用统计
被引频次:10[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://cas-ir.dicp.ac.cn/handle/321008/117968
专题中国科学院大连化学物理研究所
通讯作者Zhang LH(张丽华)
作者单位1.Chinese Acad Sci, Dalian Inst Chem Phys, Key Lab Separat Sci Analyt Chem, Natl Chromatog R&A Ctr, Dalian 116023, Peoples R China
2.Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China
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GB/T 7714
Zhu, Guijie,Zhao, Peng,Deng, Nan,et al. Single Chain Variable Fragment Displaying M13 Phage Library Functionalized Magnetic Microsphere-Based Protein Equalizer for Human Serum Protein Analysis[J]. ANALYTICAL CHEMISTRY,2012,84(18):7633-7637.
APA Zhu, Guijie.,Zhao, Peng.,Deng, Nan.,Tao, Dingyin.,Sun, Liangliang.,...&张丽华.(2012).Single Chain Variable Fragment Displaying M13 Phage Library Functionalized Magnetic Microsphere-Based Protein Equalizer for Human Serum Protein Analysis.ANALYTICAL CHEMISTRY,84(18),7633-7637.
MLA Zhu, Guijie,et al."Single Chain Variable Fragment Displaying M13 Phage Library Functionalized Magnetic Microsphere-Based Protein Equalizer for Human Serum Protein Analysis".ANALYTICAL CHEMISTRY 84.18(2012):7633-7637.
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