摘要
用过碘酸钠氧化法交联核糖基异戊烯基嘌呤和卵清蛋白，并免疫Ｂａｌ．ｂ／ｃ小鼠．鼠脾细胞与骨髓瘤细胞融合，经ＥＬＩＳＡ筛选获得抗核糖基异戊烯基嘌呤单克隆抗体．将活化的Ｓｅｐｈａｒｏｓｅ６Ｂ与单克隆抗体交联制成免疫亲和色谱柱．发酵液经免疫亲和柱纯化后用高效液相色谱检测，其纯度极高．比较了不同洗脱条件对免疫亲和柱纯化的影响，认为利用免疫亲和柱纯化发酵液具有内源性杂质干扰少的优点，是生物样品预处理的一种有效方法．
Isopentenylpurine riboside was conjugated to ovalbumin and BSA to prepare the antigen and immunize in Bal.b/c mice with IPROA. The spleen cells from an immunized mouse were fused with myleoma cells. McAb antiIPR was prepared from ascites fluid. It was coupled with Sepharose 6B to prepare the immune IgGaffinity column. The fermentation liquid containing IPR was purified with IAC column and checked by HPLC . The results showed that the IAC column had the characteristic of high purify and high specificity.
基金
浙江省自然科学基金
关键词
免疫亲和色谱
单克隆抗体
高效液相色谱
发酵液
immune affinity column
monoclonal antibody
high performance liquid chromatography
isopentenylpurine riboside
fermentation liquid
