大肠癌相关新基因ST13单克隆抗体的制备及生物学特性鉴定

Preparation and identification of the monoclonai antibody against colorectal-cancer-related gene ST13

目的 研制抗 ST13蛋白单克隆抗体 ,建立检测方法 ,探讨 ST13基因的生物学特性。方法 按淋巴细胞杂交瘤技术制备抗 ST13蛋白单克隆抗体 ,并以亲和层析法纯化单抗 ,将此单抗作探针用 Western- Blot和免疫组化检测标本。结果 获得 3个抗 ST13蛋白单抗的杂交瘤细胞株 ,杂交瘤细胞培养上清和腹水中单抗的 EL ISA效价为 10 4～ 10 5。以 ST13单抗作 Westerb- Blot和免疫组化证实该单抗具有高度特异性 ,可用于检测组织表达的ST13蛋白。结论 制备了具有高度特异性和高效价的抗 ST13蛋白单抗 ;建立了 Western- Blot及免疫组化检测组织表达的 ST13蛋白的方法。

Objective To prepare monoclonal antibody against ST13 protein and to establish detection methods for clinical application. Methods BALB/C mice were immunized with GST-ST13 fusion protein for the preparation of monoclonal antibody by hybridoma technique,and the antibody was purified by ST13 protein affinity chromatography. With the antibody some clinical specimens were exammed by Western-Blot and immunocytochemical staining. Results Three strains of hybridoma were obtained with antibody titer from 10 4 to 10 5 in tissue culture supernatant and ascites fluid. Western-Blot confirmed that the antibody specifically recognized the 50 KD ST13 protein in tissue total proteins. Using the antibody,immunocytochemical staining showed that ST13 protein was located in the cytoplasm of intestinal epithelial cell.Conclusion ST13 monoclonal antibody prepared with high specificity and high titer is useful for detection of ST13 protein in tissue specimens.\;