抗人雌激素受体单抗的制备及鉴定

DEVELOPMENT AND CHARACTERIZATION OF MONOCLONAL ANTIBODIES TO HUMAN ESTROGEN RECEPTORS

分别采用部份纯化的人乳腺癌雌激素受体（ｈＥＲ）及合成的与ｈＥＲ分子中ＤＮＡ结合区序列相同的多肽（ＡＴ３）作免疫原。经用Ｋｏｈｌｅｒ和Ｍｉｌｉｓｔｅｉｎ杂交瘤技术，制备出７株抗人雌激素受体单体（１Ｈ１２、２Ｄ７、ＡＴ３～２７等），并经ｓｅｐｈａｃｒｙｌ凝胶层析、免疫沉淀反应和间接免疫荧光标记法作特异性鉴定。其中一株高亲和力、高特异性的单抗（１Ｈ１２）在免疫沉淀反应中与人乳腺癌雌激素受体的特异性结合率达４６．２±２２．０％，并用该单抗成功地进行了人乳腺癌组织冰冻切片的ｈＥＲ间接免疫荧光标记。

Two antigens have been used to prepare monoclonal antibodies(McAbs)to human estrogen receptor (hER).We have partially purified the hER from human breast cancer tissue. And we have synthesized peptide(AT3)with amino acid sequences identical to those spanning amino acids 247～261 of the DNA-binding domain of the human estrogen receptor.Seven McAbs have been devel-oped by the classical hybridization technique. Of those McAbs that recognize both the native form of ER and the peptide (AT3)syn- thesized , one recognizes the native form of ER with high affinity,as demonstrated by gel filtration,immunoprecipitation and im-munofluorescence detection. In immunoprecipitation .the specific binding of the McAb (1H12)with the native form of ER in the breast cancer tissue is as high as 46.2±22.2%, These results demonstrate that the McAbs developed are specific to the native form of hER and will be useful in the detection of hER level in the breast cancer.