人粘蛋白糖链单抗的制备及在胃化生粘膜和胃肿瘤中的应用

制备抗胃肠道粘蛋白糖链单抗,并研究其抗原在小胃粘膜、化生粘膜及肿瘤中的表达,方法：将肠道粘膜冻干粉碎,ＣｓＣｌ平均密度离心法提取人胃肠粘蛋白,免疫ＢＡＬＢ／ｃ小鼠,取鼠脾细胞与ＳＰ２／Ｏ－Ａｇ１４鼠的骨髓瘤细胞融合成杂交瘤,克隆制备单克隆抗体,并应用ＬＳＡＢ法检测其抗原在胃粘膜,化生取粘膜和肿瘤中的表达。结果制备抗人胃粘蛋白单抗（ＨＧＭ７２,７５）和抗人肠粘蛋白单抗（ＨＣＭ１４,２１）均为抗粘蛋白

The preparation of monoclonal antibody against human mucin carbohydrates and its usage in the detection of mucin carbohydrates in gastric metaplasia and neoplasm

Objective:To prepare monoclonal antibody specific to gastrointestinal mucin carbohydrates and to detect the mucin carbohydrates expressed in gastric metaplasia and neoplasm.Methods:Human gastrointestinal mucin was extracted from both gastric and intestinal mucosa lyophilized and shattered by using CsCl equilibrium density gradient centrifugation.The splenic lymphocytes from BALB/c mice immunized with the extracts were fused with SP2/0Ag 14 mouse myeloma cells to produce hybridomas,from which specific monoclonal antibodies were prepared.The expression of mucin carbohydrates in gastric mucosa and neoplasm was determined by an immunohistochemical method.Results:Four of monoclonal antibodies specific to human mucin carbohydrates were prepared.Two of them were human gastric mucin carbohydrate specific and were named HGM 72 and HGM 75 respectively.Another two of them were colorectal mucin carbohydrate specific and named HCM 14 and HCM 21 respectively.The HGM 72 specific mucin carbohydrates could be detected in the cells of antral gland and mucosa neck cells of gastric gland.The HGM 75 specific mucin carbohydrates were demonstrated exiting in the gastric foreolar epithelial cells.Both HCM 14 and HCM 21 specific mucin carbohydrates were found in gastric metaplasia mucosa.In the tumor tissues assayed,mucin carbohydrates specific to four of the monoclonal antibodies could be detected in a lower amount compared with that in normal gastric mucosal tissues.Conclusion:Four of prepared monoclonal antibodies recognizing different epitopes of human gastrointestinal mucin carbohydrates might be used as tools to detect the expression of different mucin carbohydrates,and therefore to determine the precursor cells from which tumor cells originated.