应用HiTrap protein-A层析柱纯化新城疫病毒单克隆抗体

Purification of monoclonal antibodies of NDV using HiTrap protein-A affinity chromatography

两株针对新城疫病毒HN基因的单抗1E5和C3一B7，均具有血凝抑制性（HI），免疫球蛋白亚类分别为IgG2a和IgG1。用HiTrap protein—A层析柱对这两株单抗进行了亲和纯化。收集不同洗脱阶段的液体进行蛋白含量测定，结果表明，5mL的洗脱液洗脱后，第1～2mL洗脱液中单抗含量最高。用SDS—PAGE电泳对纯化产物进行纯度鉴定，结果表明单抗纯化后只有重链和轻链两条带。

The monoolonal antibodies 1E5 and C3-B7 ,belonging to IgG2a and IgG1 , with HI function and directed against HN gene of NDV, were purified by using affinity chromatography with HiTrap protein-A. The protein content in purge liquid was determined. The result showed that the first 1 - 2 mL elution buffer had more protein after washing with 5 mL purge liquid. The results of determination of purifica- tion McAbs by SDS-PAGE respectively indicated that every purified McAb only took on two bands in SDS-PAGE electrophoresis.