Author/Authors :
zhang, liding Research Center of Molecular Medicine of Yunnan Province - Faculty of Life Science and Technology - Kunming University of Science and Technology, China , chen, congjie Research Center of Molecular Medicine of Yunnan Province - Faculty of Life Science and Technology - Kunming University of Science and Technology, China , chen, zhixin Research Center of Molecular Medicine of Yunnan Province - Faculty of Life Science and Technology - Kunming University of Science and Technology, China , he, shuzhen Research Center of Molecular Medicine of Yunnan Province - Faculty of Life Science and Technology - Kunming University of Science and Technology, China , song, yuzhu Research Center of Molecular Medicine of Yunnan Province - Faculty of Life Science and Technology - Kunming University of Science and Technology, China , xia, xueshan Research Center of Molecular Medicine of Yunnan Province - Faculty of Life Science and Technology - Kunming University of Science and Technology, China , han, qinqin Research Center of Molecular Medicine of Yunnan Province - Faculty of Life Science and Technology - Kunming University of Science and Technology, China , zhang, jinyang Research Center of Molecular Medicine of Yunnan Province - Faculty of Life Science and Technology - Kunming University of Science and Technology, China
Abstract :
background:zika virus (zikv) is a new type of flaviviruses transmitted by mosquitoes to cause severe diseases including guillainbarré syndrome and congenital malformations during recent outbreaks. the early diagnosis of zikv infection and efficient vaccines would be effective in controlling epidemics and timely treatment. however, the native nonstructural protein 1 (ns1) mainly acquired from cells infected with zikv, which largely limits its application.objectives:this study aimed to express and purify the recombinant ns1 protein and prepare its antibody that efficiently reacted with the native ns1 protein secreted in the supernatant of zikv infected hostcells.methods:we constructed a prokaryotic expression vector containing the full length of the ns1 gene. thus, the recombinant ns1 protein was efficiently expressed and purified. the purified ns1 protein was used to prepare an antibody. the enzymelinked immunosorbent assay (elisa), western blot, and indirect immunofluorescent assay (ifa) were used to assess the reactivity and specificity of the antibody.results:the recombinant ns1 protein from a prokaryotic expression vector had good immunogenicity and the prepared antibody could specifically react with the native ns1 produced in vero, bhk21 cells infected with zikv. besides, the elisa and western blot showed that the native zikvns1 protein was secreted extracellularly and could play the role of an early diagnostic biomarker for zikv infection.conclusions:the prepared antibody against the recombinant ns1 protein is a reliable biological tool that enables the rapid and sensitive detection of secreted ns1 from host cells infected with zikv. the recombinant ns1 protein and its antibody can contribute to developing vaccines and antibodies targeting the ns1 protein to prevent flavivirus disease progression.
Keywords :
NS1 Protein , Zika Virus , Prokaryotic Expression , Antibody
