黑线毛足鼠和金仓鼠血清IgG的纯化及抗血清的制备

Purification of IgG from Phodopus sungorus and Mesocricetus auratus Sera and the Production of the Antisera

目的纯化黑线毛足鼠和金仓鼠血清IgG,制备兔抗金仓鼠和黑线毛足鼠血清IgG的抗血清。方法用Hitrap Protein G亲和层析纯化黑线毛足鼠和金仓鼠血清IgG,经SDS-PAGE电泳鉴定纯度,标准免疫方法免疫兔子制备抗血清。结果黑线毛足鼠和金仓鼠血清IgG对protein G有很高的亲合性,用Hitrap Protein G亲和层析纯化,得到高纯度黑线毛足鼠和金仓鼠IgG,利用纯化的IgG作抗原制备了高效价的抗血清,免疫双扩散测定效价达1∶32和1∶16。结论证实黑线毛足鼠和金仓鼠IgG和Protein G具有很高的亲和性,Protein G亲和层析是纯化黑线毛足鼠和金仓鼠IgG有效的方法之一,制备了黑线毛足鼠和金仓鼠IgG的抗血清。

Objective To purify IgG from Phodopus sungorus and Mesocricetus auratus and produce polyclonal rabbit antiserum against Phodopus sungorus and Mesocricetus auratus IgG. Methods The IgG of Phodopus sungorus and Mesocricetus auratus was purified by Hitrap Protein-G Affinity Chromatography. The purity of the IgG was identified by SDS-PAGE and then the IgG was subjected to immulize the rabbits to produce the antiserum. Results Both the Phodopus sangorus and Mesocricetus auratus IgG was showed high binding activity with protein-G. The IgG from Phodopus sungorus and Mesocricetus auratus was purified using Protein-G Affinity Chromatography method and the purified IgG was showed high purity on SDS-PAGE. The polyclonal rabbit antiserum against Phodopus sungorus and Mesocricetus auratus was produced by immunization of the rabbit and the titers of the antisera were up to 1 ： 32 and 1 ： 16 respetively on Double Immunodiffusions. Conclusion The method was established to purify IgG fraction for Phodopus sungorus and Mesocricetus auratus serum and the polyclonal rabbit antiserum was obtained. Our results also suggested that Phodopus sungorus and Mesocricetus auratus IgG has high binding activity with Protein G.