Broadly cross-reactive mimotope of hypervariable region 1 of hepatitis C virus derived from DNA shuffling and screened by phage display library

	Broadly cross-reactive mimotope of hypervariable region 1 of hepatitis C virus derived from DNA shuffling and screened by phage display library
	Zhang, XX; Deng, Q; Zhang, SY; Liu, J; Cai, Q; Lu, ZM; Wang, Y
刊名	JOURNAL OF MEDICAL VIROLOGY
	2003
卷号	71 期号:4 页码:511-517
关键词	hepatitis C virus hypervariable region 1 DNA shuffling phage display mimotope
通讯作者	Wang, Y (reprint author), Chinese Acad Sci, State Key Lab Mol Biol, Inst Biochem & Cell Biol, Shanghai Inst Biol Sci, 320 Yue Yang Rd, Shanghai 200031, Peoples R China.,
英文摘要	The hypervariable region 1 (HVR1) is the target of neutralizing antibodies but with isolate specificity. The aim of this study was to obtain immunogenic mimotopes of HVR1, which can react broadly with different HVR1 antibodies and could be one of the candidate immunogens in an effective vaccine against HCV. Thirty-one HVR1 cDNA fragments were digested by DNase I into a pool of random fragments and reassembled by repeated cycles of annealing in the presence of DNA polymerase to their original size. The shuffled HVR1 was then inserted into the gene III phagemid vector pCANTAB-5E and displayed on the surface of the phage. Eight individual phages were selected after four rounds of biopanning against anti-HVR1. ELISA was carried out on immobilized purified phages, respectively, to detect their reactivity with a panel of sera. DNA sequences of the inserts were analyzed and compared with the consensus sequences defined by Puntoriero et al. [(1998) EMBO J 17:3521-3533]. The reactivity of the eight selected clones to the 30 sera were from 53.3 to 80%. Among these, phage 13 (ETYVSGGSAARNAYGLTSLFTVGPAQK, aa 384-410) reacted most broadly. None of the selected sequences encoded for peptides corresponded to known HVR1 from original viral isolates. The two high reactive phages had the similar amino acid sequences with the consensus, which might play a particular role in determining the frequency of reactivity. In conclusion, this study has used effectively DNA shuffling combined with phage display technology to identify broadly cross-reactive mimotopes recognized by human polyclonal antibodies. Mimotope 13 and 23 appeared to be most reactive immunologically and could be candidate immunogens. Efforts are now underway to identify their neutralizing antibodies by immunization of animals. (C) 2003 Wiley-Liss, Inc.
学科主题	Virology
类目[WOS]	Virology
关键词[WOS]	MONOCLONAL-ANTIBODIES ; NEUTRALIZING ANTIBODIES ; LIVER-TRANSPLANTATION ; FILAMENTOUS PHAGE ; HYPERIMMUNE SERUM ; NATURAL-HISTORY ; EVOLUTION ; PROTEIN ; SEQUENCE ; PEPTIDES
收录类别	SCI
语种	英语
WOS记录号	WOS:000186101900007
内容类型	期刊论文
版本	出版稿
源URL	[http://202.127.25.143/handle/331003/2271]
专题	上海生化细胞研究所_上海生科院生化细胞研究所
推荐引用方式 GB/T 7714	Zhang, XX,Deng, Q,Zhang, SY,et al. Broadly cross-reactive mimotope of hypervariable region 1 of hepatitis C virus derived from DNA shuffling and screened by phage display library[J]. JOURNAL OF MEDICAL VIROLOGY,2003,71(4):511-517.
APA	Zhang, XX.,Deng, Q.,Zhang, SY.,Liu, J.,Cai, Q.,...&Wang, Y.(2003).Broadly cross-reactive mimotope of hypervariable region 1 of hepatitis C virus derived from DNA shuffling and screened by phage display library.JOURNAL OF MEDICAL VIROLOGY,71(4),511-517.
MLA	Zhang, XX,et al."Broadly cross-reactive mimotope of hypervariable region 1 of hepatitis C virus derived from DNA shuffling and screened by phage display library".JOURNAL OF MEDICAL VIROLOGY 71.4(2003):511-517.