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鉴定 1 型扭结绉丝病毒 ORF1 编码假定衣壳蛋白上的两个新抗原表位。

Identification of two new antigen epitopes on the putative capsid protein encoded by torque teno sus virus type 1 ORF1.

机构信息

Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of the Chinese Academy of Agricultural Sciences, Harbin 150001, China.

出版信息

Vet Microbiol. 2013 Dec 27;167(3-4):372-82. doi: 10.1016/j.vetmic.2013.09.028. Epub 2013 Oct 1.

Abstract

Torque teno sus virus type 1 (TTSuV1) ORF1 is considered to encode the viral capsid (Cap) protein, which is crucial for the induction of TTSuV1-specific antibodies and protective immunity in the host. Eight monoclonal antibodies (mAbs) directed against the Cap protein were generated and biologically characterized. The immunoreactivity of the Cap protein expressed in transfected 293T cells for these mAbs was determined with an immunoperoxidase monolayer assay. The antigen epitopes of the Cap protein were mapped using these mAbs and truncated Cap proteins expressed in Escherichia coli. Fine epitope mapping was then performed with a panel of synthesized polypeptides. All the mAbs reacted with the Cap protein C fragment expressed in E. coli. One antigenic epitope of the Cap protein, which reacted with seven mAbs, had the polypeptide sequence (536)HPKYAGQGGGYTT(548), whereas another epitope recognized by the 1E9 mAb had the polypeptide sequence (549)EIGHQGITAASLR(561). It is interesting that the two new epitopes are adjacent, but mutually independent. This study should facilitate further investigation of the antigenic differences and enable the differential diagnosis of the virus.

摘要

Torque teno sus 病毒 1 型(TTSuV1)ORF1 被认为编码病毒衣壳(Cap)蛋白,该蛋白对于诱导宿主产生 TTSuV1 特异性抗体和保护性免疫至关重要。生成并生物学表征了 8 种针对 Cap 蛋白的单克隆抗体(mAbs)。使用免疫过氧化物酶单层测定法测定转染 293T 细胞中表达的 Cap 蛋白的这些 mAbs 的免疫反应性。使用这些 mAbs 和在大肠杆菌中表达的截短的 Cap 蛋白来绘制 Cap 蛋白的抗原表位图。然后使用一组合成多肽进行精细表位作图。所有 mAbs 均与在大肠杆菌中表达的 Cap 蛋白 C 片段反应。Cap 蛋白的一个抗原表位与 7 种 mAbs 反应,其多肽序列为(536)HPKYAGQGGGYTT(548),而 1E9 mAb 识别的另一个表位具有多肽序列(549)EIGHQGITAASLR(561)。有趣的是,这两个新表位相邻,但彼此独立。这项研究应有助于进一步研究抗原差异,并能够对该病毒进行鉴别诊断。

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