[1]石磊,潘雪刁,王军业,等.噬菌体随机肽库淘选肺癌细胞NCIH1299特异性结合多肽[J].中国药理学通报,2009,(03):0.
 SHI Lei,PAN Xue diao,WANG Jun ye,et al.Panning and identification of lung cancer cell line NCIH1299 specific peptide from a phage display peptide library[J].Chinese Pharmacological Bulletin,2009,(03):0.
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噬菌体随机肽库淘选肺癌细胞NCIH1299特异性结合多肽()
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《中国药理学通报》[ISSN:/CN:]

卷:
期数:
2009年03期
页码:
0
栏目:
论著
出版日期:
2009-03-25

文章信息/Info

Title:
Panning and identification of lung cancer cell line NCIH1299 specific peptide from a phage display peptide library
作者:
石磊1 潘雪刁1 王军业2潘琴1 巫玮1 金桂芳1房思练3侯炳忠4何兴祥5臧林泉1
1.广东药学院药科学院药理教研室,2.中山大学附属肿瘤医院,3.中山大学附属第六医院,4.中山大学附属第五医院,5.广东药学院临床医学院,广州 广东510006
Author(s):
SHI Lei1 PAN Xuediao1 WANG Junye2 PAN Qin1 WU Wei1 JIN GuifangFANG Silian3
1.Dept of Pharmacology, Guangdong Pharmaceutical University; 2.the Affiliated Tumor Hospital of Sun Yatsen University;3.the Sixth Affiliated Hospital of Sun Yatsen University; 4.the Fifth Affiliated Hospital of Sun Yatsen University;5.the Clinical Medicial School of Pharmaceutical University,Guangzhou510006,China
关键词:
噬菌体肽库多肽肺癌NCIH1299减性筛选
Keywords:
phage library peptides lung cancer NCIH1299 subtract biopanning
分类号:
R05;R 32211;R 33131
文献标志码:
A
摘要:
目的利用噬菌体展示肽库筛选出与肺癌NCIH1299细胞特异性结合的多肽,鉴定其特异性和亲和力,为寻找肺癌早期诊断和治疗标志物打下基础。方法以肺癌细胞NCIH1299为靶细胞,肺二倍体成纤维细胞MRC5为吸附细胞,在37℃下对噬菌体随机十二肽库进行三轮减性筛选,挑取单克隆ELISA初步鉴定噬菌体克隆亲和力;阳性克隆提取质粒,测序后合成多肽,鉴定多肽的亲和力及特异性。结果经ELISA初步鉴定,随机挑选的20个单克隆中,其中6号对NCIH1299亲和力最高,将其命名为Phage ZS 6。细胞免疫荧光试验进一步证明,合成的相应多肽FITCZS 6对NCIH1299具有高亲和力。结论利用噬菌体随机十二肽成功筛选出与肺癌细胞具有较高亲和力的多肽ZS 6,为肺癌的早期诊断和靶向治疗奠定基础。
Abstract:
AimTo obtain polypeptides which could specifically bind to NCIH1299 cell line from peptide libraries, and to further identify and verify these polypeptides for the screening and identifying of the novel lung cancer markers in early diagnosis and treatment of lung cancer.MethodsThe lung cancer NCIH1299 cell line was used as the antigen and MRC5 was used as control for subtraction biopanning from a phage display peptide library at 37℃. The positive and specific binding clones were identified by cellbased ELISA and immunocytochemical staining, and the identified clones was sequenced. Thus the amino acid sequence was deduced and the peptide was synthesized. The peptide was identified by immunofluorescence.ResultsThrough a cellbased ELISA, immunocytochemical staining, and immunofluorescence, the Phage ZS 6 and synthetic peptide ZS 6 were shown to specially bind to the NCIH1299.ConclusionA peptide specific binding to lung cancer cell line NCIH1299 has been selected from phage display peptide libraries. Therefore, it provides a potential tool for early diagnosis of lung cancer or targeted drug delivery in chemotherapy.

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备注/Memo

备注/Memo:
收稿日期:2008-11-25,修回日期:2009-01-11基金项目:国家自然科学基金资助项目(No 30572177);教育部重点基金资助项目(No 208105)作者简介:石磊(1985-),男,硕士,研究方向:多肽药物筛选;Tel:02039352561;Email:shilei923@yahoo.com.cn;臧林泉(1965-),男,博士,教授,硕士生导师,研究方向:蛋白质/多肽药物筛选研发,分子神经药理学,肿瘤药理学,通讯作者,Tel:02039352561,Fax:02039352027,Email:zanglq@yahoo.com
更新日期/Last Update: 2009-03-25