Construction of a Phage Display Random Decapeptide Library and Panning for Mimic Antigen Epitope of ScFv A1 Against WSSV

YUAN Li; WANG Yu-zhen; XIAO Nan; ZHANG Xiao-hua; DAI He-ping

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December 2006

YUAN Li, WANG Yu-zhen, XIAO Nan, ZHANG Xiao-hua and DAI He-ping. Construction of a Phage Display Random Decapeptide Library and Panning for Mimic Antigen Epitope of ScFv A1 Against WSSV[J]. Virologica Sinica, 2006, 21(6): 614-618.

Citation: YUAN Li, WANG Yu-zhen, XIAO Nan, ZHANG Xiao-hua, DAI He-ping. Construction of a Phage Display Random Decapeptide Library and Panning for Mimic Antigen Epitope of ScFv A1 Against WSSV .VIROLOGICA SINICA, 2006, 21(6) : 614-618.

噬菌体展示随机十肽库的构建及抗WSSV单链抗体A1的模拟抗原表位的淘选和鉴定*

王玉珍 ^1,2 ,
张晓华 ^1,2 ,
戴和平 ^1,2 ,

1.
1.中国科学院水生生物研究所，湖北武汉 430072
2.
中国科学院研究生院，北京 100039

摘要

本实验以pCANTAB 5 E噬菌粒为载体，成功构建了较高容量的噬菌体展示随机十肽库，并将其应用于抗原模拟表位的淘选和鉴定。将一种特异识别对虾白斑综合症病毒（White spot syndrome virus, WSSV）的单链抗体A1对十肽库和十五肽库分别进行淘选，结果得到一系列能与单链抗体A1特异性结合的阳性克隆。将这些阳性克隆所编码的多肽氨基酸序列与已知的单链抗体A1的抗原WSSV388片段氨基酸序列做比对，发现多数阳性多肽序列都与WSSV388片段序列的C端一处K••••R••R•QS的氨基酸片段相似，由此推论单链抗体A1的模拟抗原表位可能是由该不连续氨基酸片段所构成的构象表位，而非线性表位。研究结果表明，噬菌体展示随机肽库技术是一种用于研究抗原表位结构的有效方法，有助于进一步探讨WSSV的结构蛋白的构象及功能，以及相应单链抗体与细胞受体相互作用的机理。
- 对虾白斑综合症病毒（WSSV)
- , 噬菌体展示随机肽库
- , 单链抗体（scFv）
- , 淘选
- , 模拟抗原表位

Construction of a Phage Display Random Decapeptide Library and Panning for Mimic Antigen Epitope of ScFv A1 Against WSSV

YUAN Li ^1,2 ,
WANG Yu-zhen ^1,2 ,
XIAO Nan ^1,2 ,
ZHANG Xiao-hua ¹ ,
DAI He-ping ^1*,,

1.
1.Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China
2.
Graduate School of the Chinese Academy of Sciences，Beijing 100039, China

Corresponding author: DAI He-ping,

Abstract

A phage display random decapeptide library with high titer was constructed using pCANTAB 5 E as vector and used to select and identify antigen epitopes. ScFv A1, which specifically binds the shrimp’s White spot syndrome virus (WSSV), was used to pan against the decapeptide library and a random 15mer library, respectively. A series of positive clones that bind specifically to scFv A1 were obtained. The amino acid sequences of the positive peptides were compared with WSSV388 fragment, which is the antigen of scFv A1. We found that most of the sequences of the positive peptides were similar to a fragment，K••••R••R•Q, located on C-terminal of WSSV388 fragment. We deduced that the mimic antigen epitopes of scFv A1 were conformational epitopes structured by the discontinuous amino acid fragment, but not by a linear amino acid fragment. These results show that phage display peptide library technology is a powerful method for analyzing the structure of antigen epitopes, which is helpful to further explore conformation and function of the structural proteins of WSSV, and the mechanisms of interactions of the virus with the antiviral antibodies and host cells.
- White spot syndrome virus (WSSV)
- , Phage display random peptide library
- , Single chian Fragment variable (scFv)
- , Panning
- , Mimic antigen epitope

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Construction of a Phage Display Random Decapeptide Library and Panning for Mimic Antigen Epitope of ScFv A1 Against WSSV

Corresponding author: DAI He-ping,

1. 1.Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China
2. Graduate School of the Chinese Academy of Sciences，Beijing 100039, China

Abstract: A phage display random decapeptide library with high titer was constructed using pCANTAB 5 E as vector and used to select and identify antigen epitopes. ScFv A1, which specifically binds the shrimp’s White spot syndrome virus (WSSV), was used to pan against the decapeptide library and a random 15mer library, respectively. A series of positive clones that bind specifically to scFv A1 were obtained. The amino acid sequences of the positive peptides were compared with WSSV388 fragment, which is the antigen of scFv A1. We found that most of the sequences of the positive peptides were similar to a fragment，K••••R••R•Q, located on C-terminal of WSSV388 fragment. We deduced that the mimic antigen epitopes of scFv A1 were conformational epitopes structured by the discontinuous amino acid fragment, but not by a linear amino acid fragment. These results show that phage display peptide library technology is a powerful method for analyzing the structure of antigen epitopes, which is helpful to further explore conformation and function of the structural proteins of WSSV, and the mechanisms of interactions of the virus with the antiviral antibodies and host cells.