Zang Mingxin, Li Jiaxuan, Xie Shuangyu, Cui Wen, Jiang Yanping, Xu Yigang, Qiao Xinyuan, Wang Li, Zhou Han, Liu Min, Li Yijing, Tang Lijie
College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, Heilongjiang, China.
Sheng Wu Gong Cheng Xue Bao. 2017 Aug 25;33(8):1244-1252. doi: 10.13345/j.cjb.170120.
To confirm the B cell epitope recognized by monoclonal antibody (MAb) 3G11 of bluetongue virus type 8 (BTV-8) VP2 protein prepared in our laboratory, antigen epitopes recognized by 3G11 were screened and identified by phage display technology. KLLAT sequence was found by sequencing of blue spot after four rounds panning and 283LL284 of common short peptide sequence was obtained after comparison to amino acid sequence of BTV-8 VP2 protein. The peptide sequences KLLAA, KALAT, KLAAT and KLLAT were synthesized and identified by indirect ELISA. KLLAA and KLLAT bound strongly with supernatant and as cites of 3G11 cells and reacted specifically with BTV-8 positive standard sera. Further sequence analysis showed that amino acid sequence 283LL284 was conserved among different serotypes of BTV-8 strains, and283LL284 was the key amino acids of antigen epitopes recognized by 3G11. This study laid the foundation to establish type 8 BTV specific immunological detection methods.
为了确定本实验室制备的蓝舌病病毒8型（BTV-8）VP2蛋白单克隆抗体（MAb）3G11所识别的B细胞表位，采用噬菌体展示技术筛选并鉴定3G11所识别的抗原表位。经过四轮淘选后对蓝斑进行测序，发现KLLAT序列，与BTV-8 VP2蛋白的氨基酸序列比对后得到常见短肽序列283LL284。合成肽序列KLLAA、KALAT、KLAAT和KLLAT，并通过间接ELISA进行鉴定。KLLAA和KLLAT与3G11细胞的上清液和抗原位点强烈结合，并与BTV-8阳性标准血清发生特异性反应。进一步的序列分析表明，氨基酸序列283LL284在不同血清型的BTV-8毒株中保守，且283LL284是3G11识别的抗原表位的关键氨基酸。本研究为建立BTV 8型特异性免疫检测方法奠定了基础。
