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首页> 外文期刊>Biotechnology Progress >Genome-Wide Analysis of Mouse Myeloma Cell Lines Expressing Therapeutic Antibodies
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Genome-Wide Analysis of Mouse Myeloma Cell Lines Expressing Therapeutic Antibodies

机译：表达治疗性抗体的小鼠骨髓瘤细胞系的全基因组分析

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Manufacturing cell line development involves transfection of therapeutic antibody genes into host cell lines and isolation of primary transfectomas that upon subcloning yield high expressing cell lines secreting the desired antibody.In an attempt to increase productivity of these cell lines,we set out to identify cellular genes whose expression level may affect antibody productivity.For this purpose,three different sets of mouse myeloma production cell lines expressing variable levels of three different therapeutic antibodies were subjected to microarray analysis using Murine GeneChip MG_U74Av2 arrays.A total of 456 genes were identified showing significant differential expression between at least one high expresser versus the control or its corresponding low expresser.Among these,161 genes were common among at least one set of cell lines,and 26 genes were common among two or more sets of cell lines.Functional classification revealed that a majority of these genes have biological process function related to cell metabolism and cell growth.A subset of the 26 genes that were identified as commonly regulated among any two or all three sets of cell lines were selected (by several criteria) for quantitative PCR confirmation of the microarray methodology.The expression level of two genes,Secretory Leukocyte Protease Inhibitor (SLPI) and Cell Division Cycle-6 (Cdc6),correlated with antibody productivity in at least two sets of cell lines,suggesting that they can potentially be utilized as targets for engineering a superior transfection host cell line.Additionally,these genes may be used for screening murine myeloma production cell lines for superior productivity.
机译：生产细胞系的开发涉及将治疗性抗体基因转染到宿主细胞系中，并分离原代转染瘤，亚克隆后可产生分泌所需抗体的高表达细胞系。为提高这些细胞系的生产力，我们着手鉴定细胞基因为此，使用Murine GeneChip MG_U74Av2阵列对表达三种不同治疗抗体水平可变的三组不同的小鼠骨髓瘤生产细胞系进行了微阵列分析。共鉴定出456个基因，显示出显着的差异其中至少有161个基因在至少一组细胞系中共有，而26个基因在两组或更多组细胞系中共有。这些基因中的大多数具有生物学过程与细胞代谢和细胞生长有关的ss功能。从两个或所有三组细胞系中选择被鉴定为共同调控的26个基因中的一个子集（通过几种标准），以定量PCR确认微阵列方法。至少两套细胞系中的分泌白细胞蛋白酶抑制剂（SLPI）和细胞分裂周期6（Cdc6）这两个基因的水平与抗体生产率相关，这表明它们有可能被用作工程化高级转染宿主的靶标另外，这些基因可用于筛选鼠骨髓瘤生产细胞系，以提高生产效率。

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