摘要
先经过饱和硫酸铵沉淀分离,再用Protein G亲和层析柱纯化,从马铃薯卷叶病毒(PLRV)重组CP作抗原制备的抗血清中获得了高纯度多克隆抗体(IgG),并用戊二醛法一步法进行碱性磷酸酶的标记获得了酶标抗体(IgG-AP)。将纯化的抗体与酶标抗体用于感染PLRV病叶的检测,DAS-ELISA反应呈阳性,结果表明,用重组CP制备的多克隆抗体可成功地用于PLRV的DAS-ELISA检测。本研究为PLRV重组CP多克隆抗体的大量制备及ELISA检测奠定了基础。
Polyclonal antibody（IgG） was isolated from antiserum against the recombinant coat proitein（CP） of PLRV by the treatment of saturated ammmonium sulfate,then crude IgG was purified with protein G affinity chromatography column.High purity IgG was achieved and labelled with alkaline phosphatase by one-step method of glutaraldehyde,and enzyme-conjugated IgG（IgG-AP） was obtained.When PLRV-infected potato leaf was detected by DAS-ELISA procedure using the prepared IgG and IgG-AP,positive reaction was observed,and there was no positive reaction in the detection of healthy leaf.The result indicated that the IgG against the recombinant CP and the resulting IgG-AP were successfully used in DAS-ELISA detection of potato leafroll virus.
出处
《华北农学报》
CSCD
北大核心
2011年第6期85-88,共4页
Acta Agriculturae Boreali-Sinica
基金
山东省自然基金项目(Y2008D43)
