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学科主题: 物理化学
题名: Single Chain Variable Fragment Displaying M13 Phage Library Functionalized Magnetic Microsphere-Based Protein Equalizer for Human Serum Protein Analysis
作者: Zhu, Guijie1;  Zhao, Peng1;  Deng, Nan1, 2;  Tao, Dingyin1;  Sun, Liangliang1;  Liang, Zhen1;  Zhang, Lihua1;  Zhang, Yukui1
通讯作者: 张丽华
刊名: ANALYTICAL CHEMISTRY
发表日期: 2012-09-18
DOI: 10.1021/ac3017746
卷: 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
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.
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
语种: 英语
WOS记录号: WOS:000308829700009
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/117968
Appears in Collections:中国科学院大连化学物理研究所_期刊论文

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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

Recommended Citation:
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.
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