Screening and Identification of Specific Binding Polypeptide for Gastric Cancer by A Ph.D-12　TM Phage Display Peptide Library_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

YU Mingjun , SUN Xuejun , LU Shaoying , ZHANG Wenjie.

Department of General Surgery, The First Affiliated Hospital, Medical College of Xi’an Jiaotong University, Xi’an 710061, ChinaCorresponding Author： LU Shao-ying, E-mail: robertlu@mail.xjtu.edu.cn;

Corresponding author：

Keywords：

Display peptide library; Polypeptide; Gastric cancer; BGC823 cell; Subtraction biopanning

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Abstract Full text Figures/Tables Video References Cited by

Objective 　To obtain the polypeptides specifically binding to gastric cancer BGC823 cell from 　a Ph.D-12　TM phage display peptide library (PDPL), to search the markers of gastric cancer for early diagnosis and treatment.
Methods 　The gastric cancer BGC823 cell was used as the antigen and the immortalized gastric epithelial GES cell was used as control for 3 rounds subtraction biopanning from PDPL at room temperature. The positive and specific binding clones were identified by cell enzyme-linked immunosorbent assay (ELISA) and immunochemistry staining. Those DNA sequences of identified clones were sequenced, polypeptides were marked by fluorescein FITC peptide tag, and polypeptides affinity and specificity of gastric cancer cells and tissues were identified.
Results 　After 3 rounds of panning, 20 phage clones were identified by ELISA, one of which （GC-11） was specially binding to the BGC823. Cell and tissue immunofluorescence assay further presented a high affinity of fluorescein-labeled peptide FITC-GC-11 with BGC823 and gastric cancer tissue.
Conclusion 　A peptide GC-11 which is specific binding to gastric cancer BGC823 cell and gastric cancer tissue has been selected from PDPL.

Citation： YU Mingjun,SUN Xuejun,LU Shaoying,ZHANG Wenjie.. Screening and Identification of Specific Binding Polypeptide for Gastric Cancer by A Ph.D-12　TM Phage Display Peptide Library. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2010, 17(10): 1050-1055. doi: Copy

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25.	Paschke M. Phage display systems and their applications [J]. Appl Microliol Biotechnol, 2006; 70(1): 2-11.

1. 　Smith GP, Scott JK. Libraries of peptides and proteins displayed on filamentous phage [J]. Methods Enzymol, 1993; 217(2): 228-257.
2. 　Houimel M, Dellagi K. Isolation and characterization of human neutralizing antibodies to rabies virus derived from a recombinant immune antibody library [J]. Virol Met, 2009; 161(2): 205-215.
3. 　吴懿娜, 杜欣军, 张伟伟, 等. 噬菌体抗体库技术及其应用 [J]. 细胞与分子免疫学杂志, 2010; 26(4): 404-406.
4. 　Larralde OG, Martinez R, Camacho F, et al. Identification of hepatitis A virus mimotopes by phage display, antigenicity and immunogenicity [J]. J Virol Methods, 2007; 140(1-2): 49-58.
5. 　Du B, Qian M, Zhou Z, et al. In vitro panning of a targeting peptide to hepatocarcinoma from a phage display peptide library [J]. Biochem Biophys Res Commun, 2006; 342(3): 956-962.
6. 　Jemal A, Siegel R, Ward E, et al. Cancer statistics [J]. CA Cancer J Clin, 2008; 58(2): 71-96.
7. 　刘春, 徐亮, 韩海彬, 等. 血清蛋白指纹图谱筛选胃癌血清标志物的初步探讨 [J]. 中国普外基础与临床杂志, 2009; 16(1): 23-27.
8. 　Jain RK. The next frontier of molecular medicine: delivery of therapeutics [J]. Nat Med, 1998; 4(6): 655-657.
9. 　Samoylova TI, Morrison NE, Globa LP, et al. Peptide phage display: opportunities for development of personalized anti-cancer strategies [J]. Anticancer Agents Med Chem, 2006; 6(1): 9-17.
10. 　Zhang BH, Zhang YQ, Wang JW, et al. Screening and identification of a targeting peptide to hepatocarcinoma from a phage display peptide library [J]. Mol Med, 2007; 13(526): 246-254.
11. 　Brown KC. New approaches for cell specific targeting: identification of cell selective peptides from combinatorial libraries [J]. Curr Opin Chem Biol, 2000; 4(1): 16-21.
12. 　Oyama T, Rombel IT, Samli KN, et al. Isolation of multiple cell binding ligands from different phage displayed peptide libraries [J]. Biosens Bioelec, 2006; 21(10): 1867-1875.
13. 　Shukla GS, Krag DN. Phage display selection for cell specific ligands: development of a screening procedure suitable for small tumor specimens [J]. Drug Target, 2005; 13(1): 　7-18.
14. 　Krag DN, Shula GS, Shen GP, et al. Selection of tumor-binding ligands in cancer patients with phage display libraries [J]. Cancer Res, 2006; 66(15): 7724-7733.
15. 　Lu S, Wang TD. In vivo cancer biomarkers of esophageal neoplasia [J]. Cancer Biomark, 2008; 4(6): 341-350.
16. 　Kehoe JW, Kay BK. Filamentous phage display in the new millennium [J]. Chem Rev, 2005; 105(11): 4056-4072.
17. 　Hui X, Han Y, Liang S, et al. Specific targeting of the vasculature of gastric cancer by a new tumor-homig peptide CGNSNPKSC [J]. Control Release, 2008; 131(2): 86-93.
18. 　Valadon P, Garnett JD, Testa JE, et al. Screening phage display libraries for organ-specific vascular immunotargeting in vivo [J]. Proc Natl Acad Sci, 2006; 103(2): 407-412.
19. 　Chen B, Cao S, Zhang Y, et al. A novel peptide (GX1) homing to gastric cancer vasculature inhibits angiogenesis and cooperates with TNF alpha in anti-tumor therapy [J]. BMC Cell Biol, 2009; 10(9): 63-72.
20. 　Bartlett GJ, Todd AE, Thomton JM. Inferring protein function from structure [J]. Methods Biochem Anal, 2003; 44(1): 387-407.
21. 　Watson JD, Todd AE, Bray J, et al. Target selection and determination in structural genomics [J]. Iubmb Life, 2003; 55(4-5): 249-255.
22. 　魏学明, 顾国利, 任力, 等. 胃癌组织中EGFR、C-erbB-2、VEGF及COX-2的表达及意义 [J]. 中国普外基础与临床杂志, 2009; 16(11): 900-905.
23. 　欧希龙, 关云艳, 颜芳, 等. 血管内皮生长因子165基因对人胃癌细胞凋亡的影响及机制 [J]. 世界华人消化杂志, 2008; 16(3): 307-310.
24. 　Shibuya M, Claesson-Welsh L. Signal transduction by VEGF receptors in regulation of angiogenesis and lymphangiogenesis [J]. Exp Cell Res, 2006; 312(5): 549-560.
25. 　Paschke M. Phage display systems and their applications [J]. Appl Microliol Biotechnol, 2006; 70(1): 2-11.

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Screening and Identification of Specific Binding Polypeptide for Gastric Cancer by A Ph.D-12　TM Phage Display Peptide Library

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CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Screening and Identification of Specific Binding Polypeptide for Gastric Cancer by A Ph.D-12 TM Phage Display Peptide Library

Abstract Full text Figures/Tables Video References Cited by

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Screening and Identification of Specific Binding Polypeptide for Gastric Cancer by A Ph.D-12　TM Phage Display Peptide Library