单克隆抗体的制备及鸡白细胞介素2的纯化

通过将鸡白细胞介素2(ChIL-2)基因克隆至表达载体pQE30，此蛋白在大肠杆菌中得到表达并被纯化。用纯化ChIL-2作为抗原免疫BALB/C鼠，得到了抗ChIL-2单克隆抗体。以单克隆抗体制备免疫亲和层析柱，作者首次从植物血凝素刺激的鸡淋巴细胞上清液中高度纯化了鸡白细胞介素2天然蛋白。纯化蛋白在SDS-PAGE上呈现分子质量分别为16ku和14ku的两条带，提示鸡白细胞介素2可能为糖基化蛋白。

Monoclonal Antibody Production and Purification of Native Chicken Interleukin-2

Chicken interleukin-2 (ChIL-2) gene was clo ned into a pQE30 vector, the protein was expressed in E. coli and purifi ed. A monoclonal antibody (mAb) specific to ChIL-2 was produced by immunizing B ALB/C mice with E. coli expressed ChIL-2 as an antigen. Native ChIL-2 was first isolated to high purity from the supernatant of phytohemagglutinin (PHA)- stimulated chicken lymphocytes using an immunoaffinity column prepared by coupli ng ChIL-2 mAb with Hitrap TM NHS activated Sepharose column. Purified nati ve ChIL-2 revealed two main bands corresponding to molecular masses of approxim ately16 ku and 14 ku respectively on SDS-PAGE, suggesting ChIL-2 is a glycosyl ated protein.