从噬菌体随机肽库中筛选SARS病毒N蛋白表位模拟肽的研究

Screening of SARS-CoV N Protein Mimotopes from Phage Display Peptide Library

目的获得具有生物学活性的SARSN蛋白的模拟肽。方法以抗SARS病毒N蛋白的单克隆抗体作为固相筛选分子,免疫筛选噬菌体随机十二肽库,并采用夹心ELISA、竞争抑制试验鉴定阳性克隆。结果经噬菌体富集后,从随机筛选的各46个克隆中得到N蛋白的2个不同表位,即由ELISA鉴定出单克隆抗体C00835个阳性克隆,确定氨基酸序列GXLXTPXXXXGT为SARSN蛋白的一个表位;单克隆抗体C03941个阳性克隆,确定氨基酸序列TTLPXXXXAXXX为SARS病毒N蛋白的另一个表位。结论用噬菌体随机12肽库成功筛选得到SARSN蛋白的2个不同模拟表位,为用SARS表位去探索其病毒的结构及疫苗的研制创造了条件。

Objective To obtain the bioactive mimicry peptide of SAILS N protein. Methods Monoclonal antibodies of SARS-CoV N protein was used to screen the phage display random 12-peptides library. The positive clones were identified by sandwhih ELISA and competitive ELISA. Results Positive clones were obtained from the screening. Two different mimicry epitopes were confirmed by DNA sequencing as mimotope of SARS N protein： GXLXTPXXXXGT, and TTLPXXXXAXXX. Conclution SAILS N protein mimotopes were obtained from the screening of a 12-mer phage peptide library. The results provide a new approach for SAILS viral therapy and vaccine development.