In vitro comparison of the antigen-binding and stability properties of the various molecular forms of IgA antibodies assembled and produced in CHO cells
In vitro comparison of the antigen-binding and stability properties of the various molecular forms of IgA antibodies assembled and produced in CHO cells
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DOI：
10.1073/pnas.96.6.3029
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发表时间：
1999-03-16
影响因子：
11.1
通讯作者：
Corthésy, B
中科院分区：
文献类型：
--
作者：
Berdoz, J;Blanc, CT;Corthésy, B
粘膜免疫反应的标志是在外部分泌物中产生抗原特异性分泌型 IgA (S-IgA) 抗体。 S-IgA 由两种不同细胞谱系产生的十种多肽组成，浆细胞中的重链和轻链组装成 IgA，IgA 与 J 链结合发生聚合，而分泌成分 (SC) 在穿过上皮的运输过程中添加。重组嵌合小鼠-人单体、二聚体和 S-IgA 抗体已在单个 CHO 细胞中产生，该细胞依次用携带嵌合重链和轻链、人 J 链和人 SC 的三个独立选择性标记的表达载体转染。各种分子形式的生化特征表明，各种多肽的组装产生了预期大小和共价的物种。所有嵌合IgA抗体保留了亲本小鼠IgA抗体的抗原结合能力。与缺乏相关 SC 的二聚体 IgA 相比，S-IgA 对富含蛋白酶的肠道洗涤液的抵抗力增强。使用最佳生产克隆，每 1 x 10(6) 细胞在 24 小时内回收最多 20 μg 重组 S-IgA。我们得出结论，用四种不同遗传元件从头编程的 CHO 细胞可以组装功能性嵌合 S-IgA。
The hallmark of a mucosal immune response is the production of antigen-specific secretory IgA (S-IgA) antibodies in external secretions. S-IgA consists of ten polypeptides produced in two different cell lineages, The heavy and light chains in plasma cells assemble into IgA, which on association with J chain become polymerized, whereas secretory component (SC) is added during transport across the epithelium. Recombinant chimeric mouse-human monomeric, dimeric, and S-IgA antibodies have been produced in a single CHO cell sequentially transfected with expression vectors carrying three independent selective markers for chimeric heavy and light chains, human J chain, and human SC, respectively. Biochemical characterization of the various molecular forms indicates that the assembly of the various polypeptides resulted in species of the expected size and covalence, All chimeric IgA antibodies retained the antigen-binding capacity of the parent mouse IgA antibody. The resistance of S-IgA to protease-rich intestinal washes was enhanced when compared with dimeric IgA lacking associated SC. Up to 20 mu g of recombinant S-IgA per 1 x 10(6) cells were recovered in 24 h with the best producing clones, We conclude that CHO cells programmed de novo with four different genetic elements can assemble functional chimeric S-IgA.