抗前列腺特异膜抗原单克隆抗体的制备及鉴定

目的 制备具有良好器官特异性的抗前列腺特异膜抗原（ＰＳＭ）单克隆抗体。方法 以前列腺癌细胞株ＬＮＣａｐ为免疫原，采用环磷酰胺免疫抑制法免疫小鼠，经细胞融合、选择性培养、筛选、克隆化等过程建立了抗ＰＳＭ的杂交瘤细胞株，并采用活细胞间接免疫荧光染色、ＥｎＶｉｓｏｎ＾ＴＭ两步法免疫组化染色、Ｗｅｓｔｅｒｎ ｂｌｏｔ对其特异性进行鉴定；双向琼脂扩散法鉴定单抗的亚类。结果 该单抗权与ＬＮＣａｐ及前列腺细胞膜

Preparation andidentification of monoclonal antibodies against PSM

Objective To establish a hybridoma cell line secreting monoclonal antibody (McAb) against prostate specific membrane antigen(PSM). Methods Cyclophosphamide was used to tolerize mice to antigens in PC 3 cells without PSM followed by immunization with similar antigens with PSM in the LNCaP cells. A murine McAb designated Ed 5 was generated and the specificity was treated by indirect immunofluorescence staining of living cells, EnVision TM immunohistochemistry and Western bloting. The subclass and type of the antibody were determined by double agar diffusion. Results The antibody Ed 5 reacts with only cell membrane of LNCaP cell line and prostate tissue,whereas it was no reaction with cells of PC 3, bladder cancer renal cancer and all the leukocytes from the 30 patients.Western blotting demonstrated that the molecules which react with Ed 5 is about 10 000.The subclass and type of the antibody belong to IgG 1 and Kappa light chain respectively. Conclusions Ed 5 reacts with PSM and this antigen is expressed on most of the prostate samples including normal prostate tissue,BPH tissues and prostate cancer.It was showed the antibody could be used in targeting diagnosis and treatment of prostate cancer.