抗人SSX2分子单克隆抗体的制备及初步应用

Preparation and characterization of monoclonal antibodies against human SSX2 molecule

目的 制备抗滑膜肉瘤X断点 2 (Synovialsarcoma,Xbreakpoint2 ,SSX2 )分子的单克隆抗体 ,探讨SSX2的组织分布及其与肿瘤的关系。方法 应用淋巴细胞杂交瘤技术制备小鼠源性抗人SSX2mAb。应用间接ELISA测定腹水效价 ,间接免疫荧光胞内染色检测抗体与肿瘤细胞系的反应性。用酶免疫组织化学技术 ,对SSX2分子在人正常睾丸组织和直肠癌组织的分布进行了鉴定。结果 获得 1株分泌抗SSX2mAb的杂交瘤细胞株LX SSX2 ,间接ELISA测定腹水效价达 1× 10 - 5,为IgG1(κ)亚类 ,能识别K5 6 2胞内天然及变性SSX2分子。在酶免疫组织化学技术应用中获得较满意效果 ,SSX2分子表达于睾丸精原细胞的细胞核内 ,并在直肠癌标本中检出到SSX2的表达。结论 LX SSX2mAb有很好的特异性 。

Objective To prepare monoclonal antibody against SSX2 molecule and to explore the distribution of SSX2 in normal human testis tissue and human rectal cancer. Methods Murine mAbs were prepared using hybridoma technique. The titer of the ascitic mAb was detected by indirect ELISA. The antibody reactivity to tumor cell line was assayed by intracellular immunofluorescence staining. The distribution of SSX2 in normal testis and rectal cancer tissue was identified by enzyme-immunohistochemical staining. Results One hybridoma cell line secreting mAb against human SSX2 molecule was obtained and named LX-SSX2. The titer of the ascitic mAb reached 10 -5 detected by indirect ELISA and the Ig subclass of LX-SSX2 was IgG1 (κ). The LX-SSX2 could react with native and denatured SSX2 molecule in K562 cells. The LX-SSX2 mAb also could be used in immunohistochemical staining. The expression of SSX2 molecule was detected in nuclear of spermatogonia as well as in rectal cancer sample. Conclusion LX-SSX2 mAb is an useful tool, which can be employed in the studies of the SSX2 distribution at protein level and the studies of the relationship between SSX2 expression and malignant tumors progressing.