抗血清
多克隆抗体
重组DNA
分子生物学
免疫印迹
亲和层析
效价
病毒学
抗体
大肠杆菌
生物
紫胶操纵子
病毒
化学
生物化学
酶
免疫学
基因
作者
Yihao Wang,Mingzhi Li,Mengle Jia,Lingdi Yang,Junjie Xiong,Ting Wang,Yu Wang,Shurong Liu,Weihong Guo,Lingbao Kong,Meifeng Li
出处
期刊:PubMed
日期:2023-07-01
卷期号:39 (7): 642-648
摘要
Objective To express the monkeypox virus (MPXV) A23R protein in Escherichia coli and purify by Ni-NTA affinity column, and to prepare mouse antiserum against MPXV A23R. Methods The recombinant plasmid pET-28a-MPXV-A23R was constructed and transformed into Escherichia coli BL21 to induce the expression of A23R protein. After optimizing the conditions of expression, A23R protein was highly expressed. Recombinant A23R protein was purified by Ni-NTA affinity column and identified by Western blot analysis. The purified protein was used to immunize mice for preparing the A23R polyclonal antibody, and the antibody titer was detected by ELISA. Results The expression of A23R recombinant protein reached the peak under the induced conditions of 0.6 mmol/L isopropyl-β-D-thiogalactoside (IPTG), 37 DegreesCelsius and 20 hours. The purity of the protein was about 96.07% and was identified by Western blot analysis. The mice were immunized with recombinant protein, and the titer of antibody reached 1:102 400 at the 6th week after immunization. Conclusion MPXV A23R is expressed highly and purified with a high purity and its antiserum from mouse is obtained with a high titre.
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