Establishment of a human somatic hybrid cell line for recombinant protein production
Establishment of a human somatic hybrid cell line for recombinant protein production
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DOI：
10.1159/000067294
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发表时间：
2002-11-01
影响因子：
11
通讯作者：
Chan, S
中科院分区：
文献类型：
--
作者：
Cho, MS;Yee, H;Chan, S
使用细胞融合技术来获得适合于大规模生产治疗性蛋白质的哺乳动物宿主细胞系。尽管人胚肾来源的293 S细胞系是哺乳动物基因表达的优良宿主细胞，但这些细胞在悬浮培养物和生物反应器中倾向于形成大而紧密的聚集体。为了解决聚集的问题，使用聚乙二醇（PEG）将293 S细胞与人悬浮细胞系21318（伯基特淋巴瘤衍生物）融合。在补充有次黄嘌呤-氨基蝶呤-胸苷和G418（1 mg/ ml）的培养基中选择PEG处理的293 S和21318细胞以消除未融合的细胞。这些杂交克隆，命名为HK B（肾和B细胞的杂交体），对内源性免疫球蛋白表达呈阴性。大多数克隆很容易适应在振荡条件下的无血清悬浮培养，而不会形成大而紧密的聚集体。在与293和中国仓鼠卵巢细胞的并行比较中，一个克隆HKB 11显示支持细胞因子[白细胞介素（IL）-2和IL-4]、ICAM-1和rFVIII的高水平表达。HKB细胞的上述特征表明，HKB 11是生产人治疗性蛋白质的有利细胞宿主。版权所有（C）2002国家科学理事会，中华民国和S. Karger AG，巴塞尔。
Cell fusion techniques were used to derive mammalian host cell lines suitable for large-scale production of therapeutic proteins. Although the 293S cell line, of human embryonic kidney origin, is an excellent host cell for mammalian gene expression, these cells have a tendency to form large and tight aggregates in suspension cultures and bioreactors. To solve the problem of aggregation, 293S cells were fused to a human suspension cell line, 21318 (a Burkitt's lymphoma derivative), using polyethylene glycol (PEG). The PEG-treated 293S and 21318 cells were selected in a medium supplemented with hypoxanthine-aminopterin-thymidine and G418 (1 mg/ ml) to eliminate nonfused cells. These hybrid clones, designated as HKB (hybrid of kidney and B cells), are negative for endogenous immunoglobulin expression. Most clones are readily adaptable to serum-free suspension culture under shaking conditions without forming large and tight aggregates. One clone, HKB11, was shown to support high-level expression of cytokines [interleukin (IL)-2 and IL-4], ICAM-1 and rFVIII in a side-by-side comparison with 293 and Chinese hamster ovary cells. The above-described characteristics of HKB cells indicate that HKB11 is a favorable cell host for the production of human therapeutic proteins. Copyright (C) 2002 National Science Council, ROC and S. Karger AG, Basel.