Development of anti-human colonic mucin monoclonal antibodies. Characterization of multiple colonic mucin species.
Development of anti-human colonic mucin monoclonal antibodies. Characterization of multiple colonic mucin species.
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抗人结肠粘蛋白单克隆抗体的开发。
DOI：
10.1172/jci112428
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发表时间：
1986
期刊：
影响因子：
--
通讯作者：
K. Lynch
中科院分区：
文献类型：
--
作者：
D. Podolsky;D. Fournier;K. Lynch
使用针对纯化的人结肠粘蛋白（HCM）的单克隆抗体（MAbs）文库评估结肠粘蛋白种类之间的结构关系。在用来自正常人结肠粘膜的纯化粘蛋白免疫小鼠后，在固相测定中筛选的1，920个融合产物中有14%的抗HCM活性呈阳性。杂交瘤的选择性结合模式，以六个离散的HCM物种（I-VI）分离的DEAE-纤维素色谱法建议HCM物种I-VI之间的共享和物种特异性抗原决定簇的存在。制备了23种抗HCM单克隆抗体（7种IgM，7种IgG 1和9种IgG 2），证明了一系列抗HCM物种特异性，并用于研究粘蛋白物种之间的结构关系。通过竞争性固相放射免疫测定法显示了各个抗HCM单克隆抗体对各种粘蛋白物种的结合，以反映不同物种上存在相同的表位。用抗HCM单克隆抗体制备的各种亲和树脂上的HCM种类的吸附表明，由单个单克隆抗体结合到多个粘蛋白种类与内在结构决定因素有关。四个抗HCM单克隆抗体识别蛋白酶敏感的抗原结构，这表明它们可能是针对核心HCM蛋白。通过固相测定显示12种抗HCM MAb识别具有确定结构的完整（n = 5）或部分（n = 7）分离的结肠粘蛋白寡糖侧链。总的来说，这些数据表明结肠粘蛋白物种之间存在共享和独特的抗原结构决定簇。粘蛋白糖蛋白的色谱异质性似乎与正常人结肠中存在的具有生物学意义的亚类有关。
Structural relationships between colonic mucin species were assessed using a library of monoclonal antibodies (MAbs) directed against purified human colonic mucin (HCM). After immunization of mice with purified mucin from normal human colonic mucosa, 14% of 1,920 fusion products screened were positive for anti-HCM activity in a solid-phase assay. Patterns of selective binding by hybridomas to six discrete HCM species (I-VI) separated by DEAE-cellulose chromatography suggested the presence of both shared and species-specific antigenic determinants among HCM species I-VI. 23 anti-HCMs MAbs (7 IgM, 7 IgG1, and 9 IgG2) demonstrating a range of anti-HCM species specificities, were produced and used to study structural relationships between mucin species. Binding of various mucin species by individual anti-HCM MAbs was shown by competitive solid-phase radioimmunoassay to reflect the presence of identical epitopes on the different species. Adsorption of HCM species on a variety of affinity resins prepared with anti-HCM MAbs demonstrated that binding to multiple mucin species by a single MAb was related to intrinsic structural determinants. Four anti-HCM MAbs recognized protease-sensitive antigenic structures, which suggests that they may be directed to core HCM proteins. 12 of the anti-HCM MAbs were shown by solid-phase assay to recognize either complete (n = 5) or partial (n = 7) isolated colonic mucin oligosaccharide side chains of defined structure. Collectively, these data show the presence of both shared and unique antigenic structural determinants among colonic mucin species. Chromatographic heterogeneity of mucin glycoproteins seems to be related to the existence of biologically significant subclasses in the normal human colon.
从大鼠结肠粘液糖蛋白中分离和表征寡糖。
DOI：
--
发表时间：
1980
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Slomiany,BL;Murty,VL;Slomiany,A
通讯作者：
Slomiany,A