Preparation and application of the rabbit anti-hamster IgG-HRP
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KeyWord:Golden-hamster; IgG; Affinity chromatography; Rabbit anti-hamster IgG-HRP
刘先菊 中国医学科学院实验动物研究所 卫生部实验动物检测中心, 北京
林树柱 中国医学科学院实验动物研究所 卫生部实验动物检测中心, 北京
杨帆 中国医学科学院实验动物研究所 卫生部实验动物检测中心, 北京
佟巍 中国医学科学院实验动物研究所 卫生部实验动物检测中心, 北京
刘云波 中国医学科学院实验动物研究所 卫生部实验动物检测中心, 北京
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      ObjectiveTo purify IgG from hamster serum, and to prepare the rabbit anti-hamster IgG-HRP, to be used for detection of the antibody from Sendai virus (SV)-infected hamsters. MethodsUsing HiTrap Protein-A affinity chromatography column to purify the IgG from hamster serum. The purity of the hamster-IgG was identified by SDS-PAGE analysis. To produce the hamster against rabbit IgG, and to measure the titer of rabbit anti-hamster IgG by double immunodiffusion. To purify the rabbit anti-hamster IgG by affinity chromatography and prepare the rabbit anti-hamster HRP-conjugated antibodies. The working concentration of the rabbit anti-hamster IgG-HRP was detected by direct ELISA and Western blot, and detect the antibody from Sendai virus-infected hamster by enzyme immunoassay (IEA). ResultsThe hamster IgG was purified up to 95% determined by SDS-PAGE analysis, and the titer of the antisrum was up to 1:64 assessed by double immunodifusion. The working concentration of the rabbit anti-hamster IgG was up to 1:5000 measured by direct ELISA, and 1:2000 measured by Western blot. The IEA titer was 1:2000. ConclusionsThe method has been established for efficient purification of IgG fraction from hamster serum, and the polyclonal antibody of the hamster-IgG and the rabbit anti-hamster IgG-HRP are prepared, which may provide a basis for development of serological test system of microbial pathogens of hamster.
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