目的 寻找识别新的肿瘤相关粘蛋白抗原的单抗,为阐明胃肠癌粘液糖蛋白的表达规律进而为胃肠肿瘤的预后、分型等提供参考依据。方法 以牛颌下腺粘蛋白(BSM)为免疫原免疫小鼠,以免疫脾细胞与Sp2/0细胞融合建立杂交瘤细胞株,用有限稀释法克隆化,间接ELISA及间接ELISA结合神经氨酸酶消化筛选克隆。以聚丙烯酰胺凝胶电泳,免疫印记(Western-Blot),间接ELISA结合热处理、胰蛋白酶消化、过碘酸氧化以及稀碱处理去除O-乙酰基等方法进行抗体的鉴定。以免疫组化LSAB法观察两株抗体对68例胃癌和55例肠癌粘蛋白糖链抗原的标记情况。结果 两株抗体分别命名为5E2和6C10,Western-Blot显示两株抗体均只与分子量大于220kDa的PAS染色阳性条带反应。两株单抗与BSM的反应都对热处理、胰蛋白酶消化、过碘酸氧化、神经氨酸酶消化等处理敏感。5E2还对稀碱去除O-乙酰基敏感,6C10可以部分吸收sialyl-lewis~a。单抗6C10在早期胃癌及癌旁的阳性率分别为5.5％和52.9％,进展癌及其癌旁的阳性率分别为14.0％和22.9％;在结肠癌及其癌旁的阳性率分别为30.9％和83.8％。单抗5E2在胃早期癌及其癌旁的阳性率分别为16.7％和41.2％,在进展癌及其癌旁的阳性率分别为42.0％和37.5％;在肠癌及其癌旁的阳性率分别为69.1％和91.9％。结论 初步认为单 Objective To search for monoclonal antibodies that recognize new tumor-associated mucin antigens to provide reference for the prognosis and classification of gastrointestinal tumors in order to elucidate the expression patterns of mucin glycoprotein in gastrointestinal cancer. Methods Bovine submandibular gland mucin (BSM) was used as immunogen to immunize mice. The spleen cells were fused with Sp2 / 0 cells to establish hybridoma cell lines. The hybridoma cells were cloned by limiting dilution, indirect ELISA and indirect ELISA with neuraminidase Screen clones. Antibodies were identified by polyacrylamide gel electrophoresis, Western-Blot, indirect ELISA with heat treatment, trypsin digestion, periodic acid oxidation and dilute alkaline treatment to remove O-acetyl. Immunohistochemistry LSAB method observed two antibodies on 68 cases of gastric cancer and 55 cases of mucinous mucosa glycogen antigen marker. Results The two antibodies were named as 5E2 and 6C10 respectively. Western-Blot showed that both antibodies only reacted with PAS staining positive bands with molecular weight greater than 220 kDa. The reaction of two McAbs with BSM was sensitive to heat treatment, trypsin digestion, periodic acid oxidation, neuraminidase digestion and other treatments. 5E2 is also sensitive to dilute alkali to remove O-acetyl, 6C10 can partially absorb sialyl-lewis ~ a. The positive rates of McAb 6C10 in early gastric cancer and paracancer were 5.5% and 52.9%, respectively, and the positive rates in advanced cancer and paracancer were 14.0% and 22.9% respectively. The positive rates in colorectal cancer and paracancerous tissues were 30.9 % And 83.8%. The positive rates of McAb 5E2 in gastric cancer and its adjacent tissues were 16.7% and 41.2% respectively, and were 42.0% and 37.5% respectively in patients with advanced cancer and its adjacent tissues. The positive rates in colorectal cancer and its adjacent tissues Respectively, 69.1% and 91.9%. Conclusion initially considered a single