一种新型胃肠道癌特异性单链抗体的表达及其在胃癌中的检测

摘要

Objective To construct a single chain fragment variable (scFv) to tumor associated antigen with gastrointestinal carcinoma, identify and check the expression of scFv in the human gastric carcinoma cells. Mothods The expression of scFv gene in soluble or secrete manners were determined by Western blot and ELISA (enzyme-linked immuneosorbent assay). After several human gastric carcinoma cells were cultured, the tumor associated antigen was determined with the scFv by immunohistochemistry in the human gastric carcinoma cells. Results It proved the scFv was a 31 KD protein. The scFv could identify and bind to gastric carcinoma cell lines KATO Ⅲ, HGC-27 and hepatocellular carcinoma cell SMMC7721. However, it could not bind to SGC7901. Conclusion Somes properties of the scFv suggests that it is possible therapeutic applications.%目的通过原核可溶性表达将我们制备的抗胃肠道肿瘤特异性单链抗体(single chain antibody variable fragment,scFv)基因表达出单链抗体蛋白,初步鉴定并检测其在几种胃癌细胞中的表达.方法通过基因原核表达、Western blot、ELISA等方法检测单链抗体的表达;通过免疫细胞化学方法检测其对几种胃癌细胞的肿瘤相关抗原的识别.结果获得的单链抗体分子量31 KD,与理论分子量基本一致,并通过Western blot、ELISA加以证实;免疫细胞化学方法证明该单链抗体可识别胃癌细胞KATOⅢ、HGC-27、肝癌细胞SMMC7721等细胞,不能识别SGC7901.结论 通过对单链抗体的原核表达,明确了单链抗体对胃癌细胞的识别能力,为进一步临床应用提供理论基础.