Identification of the gastrointestinal and pancreatic cancer-associated antigen detected by monoclonal antibody 19-9 in the sera of patients as a mucin.
Identification of the gastrointestinal and pancreatic cancer-associated antigen detected by monoclonal antibody 19-9 in the sera of patients as a mucin.
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发表时间：
1983-11
期刊：
影响因子：
11.2
通讯作者：
J. Magnani;Z. Steplewski;H. Koprowski;V. Ginsburg
中科院分区：
文献类型：
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作者：
J. Magnani;Z. Steplewski;H. Koprowski;V. Ginsburg
由用人结肠癌细胞系免疫的小鼠的脾细胞制备的杂交瘤产生的单抗19-9在大多数胃肠道和胰腺癌患者的血清中检测到抗原(M.Herlyn，H.F.Sears，Z.Steplewski和H.Koprowski，J.Clin)。免疫学，2：135-140,1982)。该抗体的表位是具有糖序列的碳水化合物(式；见正文)，其中NeuNAc是N-乙酰神经氨酸，Gal是半乳糖，GlcNAc是N-乙酰氨基葡萄糖，Fuc是岩藻糖。在结肠癌细胞系以及许多胃肠道和胰腺癌中，这个序列出现在含有唾液酸Lea活性五糖(唾液酸化的Lea-活性五糖，IV3-α-NeuNAc-III4-α-Fuc-LcOse4，其中LcOse4是Galβ1-3GlcNAc Beta 1-3Gal Beta 1-4Glc)的单唾液酸神经节苷脂中(J.L.Magnani et al.J.Biol.化学，257：14365-14369,1982年)。然而，根据以下证据，患者血清中的抗原主要以粘蛋白的形式存在，而不是神经节苷脂。用有机溶剂从血清中提取很少的抗原，在薄层层析的条件下，提取的抗原保留在原始处，神经节苷脂抗原沿平板向上移动。血清经Sephacryl S-400凝胶过滤后，抗原在空隙中洗脱，表明其相对分子质量大于或等于5×10(6)。在0.1N NaOH中37度孵育5小时可破坏血清抗原，但不影响神经节苷脂抗原。在氯化铯梯度中测定的血清抗原密度为1.50克/毫升，而在4-甲基胍中测定的血清抗原密度为1.50克/毫升。用抗体19-9从一例Le(a-b+)血型癌症患者血清中纯化的抗原亲和力中含有Leb抗原，与粘蛋白表现出的多重抗原特异性一致。
Monoclonal antibody 19-9, produced by a hybridoma prepared from spleen cells of a mouse immunized with a human colon carcinoma cell line, detects an antigen in the serum from most patients with gastrointestinal and pancreatic cancer (M. Herlyn, H.F. Sears, Z. Steplewski, and H. Koprowski, J. Clin. Immunol., 2: 135-140, 1982). The epitope of this antibody is a carbohydrate with the sugar sequence (formula; see text) in which NeuNAc is N-acetylneuraminic acid, Gal is galactose, GlcNAc is N-acetylglucosamine, and Fuc is fucose. In the colon carcinoma cell line and many gastrointestinal and pancreatic cancers, this sequence occurs in a monosialoganglioside containing a sialylated Lea-active pentasaccharide (sialylated lacto-N-fucopentaose II, IV3-alpha-NeuNAc-III4-alpha-Fuc-LcOse4, in which LcOse4 is Gal beta 1-3GlcNAc beta 1-3Gal beta 1-4Glc) (J. L. Magnani et al. J. Biol. Chem., 257: 14365-14369, 1982). However, the antigen in the sera of patients occurs mainly as a mucin, not a ganglioside, based on the following evidence. Little antigen is extracted by organic solvents from sera, and that which is extracted remains at the origin under conditions of thin-layer chromatography where the ganglioside antigen migrates up the plate. Upon gel filtration of serum on Sephacryl S-400, the antigen is eluted in the void volume, indicating a molecular weight of greater than or equal to 5 X 10(6). Incubation for 5 hr at 37 degrees in 0.1 N NaOH destroys the serum antigen but does not affect the ganglioside antigen. The density of the serum antigen as determined in a CsCl gradient is 1.50 g/ml, while in 4 M guanidine. HCl its density is 1.43 g/ml. Finally, antigen affinity purified by antibody 19-9 from the serum of a cancer patient belonging to the Le(a-b+) blood group contains Leb antigen, consistent with the multiple antigenic specificities exhibited by mucins.