摘要
以基因工程人表皮生长因子（ｒｈ Ｅ Ｇ Ｆ）为抗原，免疫 Ｂ Ａ Ｌ Ｂ／ｃ 小鼠，将免疫的小鼠脾细胞与小鼠 Ｓｐ２／０ Ａｇ１４ 骨髓瘤细胞进行融合，经多次筛选和再克隆化，获得３ 株能稳定分泌抗ｒｈ Ｅ Ｇ Ｆ单克隆抗体（ Ｍｃ Ａｂｓ）的杂交瘤细胞株。对该３ 株杂交瘤细胞小鼠腹水进行了抗体效价测定，结果显示在１×１０－ ６～２×１０－ ７ 之间。对上述３ 种杂交瘤单抗（分别命名为： Ａ Ｅ２ Ａ４ ， Ａ Ｅ２ Ｇ５ 和 Ａ Ｅ３ Ｅ１１ ）进行了特异性、抗原决定簇及相对亲和力测定。结果表明，它们都具有抗ｒｈ Ｅ Ｇ Ｆ高特异性；并分别抗两个不同抗原决定簇；相对亲和力大小次序为： Ａ Ｅ３ Ｅ１１ ＞ Ａ Ｅ２ Ａ４ ＞ Ａ Ｅ２ Ｇ５ 。 Ｉｇ 亚类鉴定结果显示： Ａ Ｅ２ Ａ４ ， Ａ Ｅ３ Ｅ１１同为 Ｉｇ Ｇ１ 亚类，而 Ａ Ｅ２ Ｇ５ 为 Ｉｇ Ｇ２ｂ 亚类。确定 Ａ Ｅ２ Ａ４ 单克隆抗体可用于制备高纯ｒｈ Ｅ Ｇ Ｆ。
BALB/c mice were immunized with rhEGF antigen,and the immunized spenocytes were fused with murine Sp2/0 Ag14 myeloma cells.After several subcloning and screening,three hybridomas secreting monoclonal antibodies(McAbs) against rhEGF stably were established.Ascites titers of the hybridomas were done and the results were obtained as follows:from 1×10 -6 to 2×10 -7 .Analyses of specificity,antigenic determinant and relative affinity of the 3 McAbs(named AE 2A 4,AE 2G 5 and AE 3E 11 )were conducted.The results reveal that they are all highly specific to rhEGF(anti-distinct two antigenic determinants),the order of relative affinity is AE 3E 11 >AE 2A 4>AE 2G 5.Ig subclasses,AE 2A 4,AE 3E 11 :IgG1,and AE 2G 5:IgG2b.The McAb of AE 2A 4 can be used in preparing the pure rhEGF.
出处
《河北省科学院学报》
CAS
1999年第3期45-49,共5页
Journal of The Hebei Academy of Sciences
基金
河北省"九五"重点科技攻关项目
