中南医学科学杂志2023，Vol.51Issue(6)：829-833,5.DOI:10.15972/j.cnki.43-1509/r.2023.06.007
SARS-CoV-2核衣壳蛋白的生物信息学分析、纯化及多克隆抗体制备
Bioinformatics analysis,purification and immunological characterization of nucleo-capsid protein of SARS-CoV-2
摘要
Abstract
Aim To analyze the biological properties,purification and polyclonal antibody preparation of nucleo-capsid(N)protein of novel coronavirus(SARS-CoV-2).Methods The amino acid sequences of the N protein coro-naviruses that can infect human beings were download from NCBI,and the sequences were aligned with Clustal Omega soft-ware.The secondary structure of SARS-CoV-2 N protein was predicted by SOPMA software.The intrinsic disorder do-main of SARS-CoV-2 N protein was predicted by Pondr and IUPred,and the molecular recognition characteristic sequence of SARS-CoV-2 N protein was predicted by MoRFchib software;IEDB website was used to predict its B cell epitope.Pu-rified N protein was used to immunize BALB/c mice,isolate mouse serum,and prepare polyclonal antibodies.Polyclonal antibody titer in serum was determined by ELISA.The proliferation of splenocytes stimulated by N protein was detected by MTT assay.Results The homology between SARS-CoV-2 N protein and SARS-CoV N protein was higher than 97%.The content of random coil is as high as55.13%.The content of α-helix was21.24%,β-sheet was 16.71 and β-turn was 6.92%.The SARS-CoV-2 N protein contains a large number of intrinsic disorder structures,with 4 molecular recog-nition characteristic sequences and 11 linear B cell epitopes.The purified N protein has a molecular weight of 51.5 kDa and high purity,which can stimulate mice to secrete high titer antibody and cause lymphocyte proliferation.Conclusion The SARS-CoV-2 N protein contains a large number of intrinsic disorder domain and the high titer polyclonal antibody was obtained.关键词
冠状病毒/核衣壳蛋白/生物信息学/免疫学性质Key words
coronavirus/nucleocapsid protein/bioinformatics/immunological properties分类
医药卫生引用本文复制引用
王巧,陈芝喜..SARS-CoV-2核衣壳蛋白的生物信息学分析、纯化及多克隆抗体制备[J].中南医学科学杂志,2023,51(6):829-833,5.基金项目
湖南省自然科学基金(2021JJ70115) （2021JJ70115）
衡阳市科技局项目(2020hcjz6717) （2020hcjz6717）
