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。
关键词:
马铃薯卷叶病毒,
重组CP,
多克隆抗体,
酶标抗体,
DAS-ELISA
Abstract: 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, a nd 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.
Key words:
Potato leafroll virus(PLRV),
Recombinant CP,
Polycolonal antibody,
Phosphatase-conjugated IgG,
DAS-ELISA
中图分类号:
李楠楠, 左玉玲, 隋炯明, 盖树鹏, 樊连梅, 李广存, 郭宝太. 重组CP多克隆抗体在马铃薯卷叶病毒DAS-ELISA检测中的应用[J]. 华北农学报, 2011, 26(6): 85-88. doi: 10.7668/hbnxb.2011.06.016.
LI Nan-nan, ZUO Yu-ling, SUI Jiong-ming, GAI Shu-peng, FAN Lian-mei, LI Guang-cun, GUO Bao-tai. Application of Polyclonal Antibody against the Recombinant CP to the DAS-ELISA Detection of Potato Leafroll Virus[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2011, 26(6): 85-88. doi: 10.7668/hbnxb.2011.06.016.