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公开(公告)号：US20100132059A1
公开(公告)日：2010-05-27
申请号：US12615710
申请日：2009-11-10
申请人： Kevin Michael MCCARTHY , Robin Allison Heller-Harrison , Mark William Leonard , Timothy Shea Charlebois
发明人： Kevin Michael MCCARTHY , Robin Allison Heller-Harrison , Mark William Leonard , Timothy Shea Charlebois
摘要： The present invention is based on the discovery that the transcriptional activity of both hamster mammary tumor 7 (HMT-7) and hamster layilin genes are increased when Chinese Hamster Ovary (CHO) cells are cultured at temperatures lower than 37° C. The present invention provides novel genomic, cDNA, and protein sequences of HMT-7 and a novel genomic sequence for hamster layilin. Included in the genomic sequences are novel temperature-induced promoter sequences for each; two promoter sequences are provided for HMT-7 and one promoter sequence is provided for layilin. The invention additionally provides antisense and siRNA molecules to the nucleic acid molecules of HMT-7. The invention also provides genetically engineered expression vectors comprising the novel temperature-induced promoters of the invention, which are particularly useful as part of a mammalian temperature-inducible expression system. The invention also provides host cells and/or transgenic animals comprising the novel nucleic acid molecules of the invention. The invention further provides methods of using the polynucleotides of the invention, e.g., for temperature-induced transgene expression.
摘要翻译： 本发明是基于以下发现：当在低于37℃的温度培养中国仓鼠卵巢（CHO）细胞时，仓鼠乳房肿瘤7（HMT-7）和仓鼠蛋白基因的转录活性增加。本发明 提供HMT-7的新型基因组，cDNA和蛋白质序列以及仓鼠铺板的新型基因组序列。 基因组序列中包含的每一个都是新的温度诱导的启动子序列; 为HMT-7提供了两个启动子序列，并为织物提供了一个启动子序列。 本发明还向HMT-7的核酸分子提供反义和siRNA分子。 本发明还提供了包含本发明的新型温度诱导启动子的基因工程表达载体，其特别可用作哺乳动物温度诱导表达系统的一部分。 本发明还提供了包含本发明的新型核酸分子的宿主细胞和/或转基因动物。 本发明进一步提供使用本发明的多核苷酸的方法，例如用于温度诱导的转基因表达。
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2.发明申请Methods For Identifying Cells Suitable For Large-Scale Production of Recombinant Proteins 审中-公开用于鉴定适合大规模生产重组蛋白的细胞的方法公开(公告)号：US20110014624A1
公开(公告)日：2011-01-20
申请号：US12921717
申请日：2009-03-12
申请人： Kevin Michael McCarthy , Megan Jill Hone , Robin Allison Heller-Harrison , Mark William Leonard
发明人： Kevin Michael McCarthy , Megan Jill Hone , Robin Allison Heller-Harrison , Mark William Leonard
CPC分类号： C12N15/85 , C07K16/00 , C07K16/2803 , C12N2830/42 , C12N2840/203
摘要： The present invention provides methods of identifying a clonal population of cells suitable for large-scale production of a protein of interest. The invention further provides methods for high-throughput screening for genetic rearrangements in the gene encoding the protein of interest, whereby the absence of a deletion in the gene encoding the protein of interest indicates that the cell is suitable for large-scale production of the protein of interest.
摘要翻译： 本发明提供了鉴定适合于大规模生产目的蛋白质的细胞的克隆群体的方法。 本发明还提供了用于在编码目的蛋白质的基因中进行遗传重排的高通量筛选的方法，由此在编码目的蛋白质的基因中缺失缺失表明该细胞适合于大规模生产蛋白质 出于兴趣。
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3.发明申请METHODs OF DETECTION AND QUANTIFICATION OF HOST CELL DNA CONTAMINATION OF PURIFIED PROTEINS 审中-公开检测和定量纯化蛋白质细胞DNA污染的方法公开(公告)号：US20090325175A1
公开(公告)日：2009-12-31
申请号：US12476101
申请日：2009-06-01
IPC分类号： C12Q1/68
CPC分类号： C12Q1/686
摘要： The present invention provides a novel robust, sensitive, reproducible, and accurate method of detecting and quantifying host cell genomic DNA contamination utilizing quantitative real time Polymerase Chain Reaction (qPCR), wherein the qPCR primers are complementary to the highly repetitive host cell genomic DNA sequences, e.g., Alu-equivalent sequences. The present invention is particularly useful for determining the levels of residual genomic DNA in biological products to be administered as therapeutics, e.g., therapeutic proteins.
摘要翻译： 本发明提供了使用定量实时聚合酶链式反应（qPCR）检测和定量宿主细胞基因组DNA污染的新颖的鲁棒，灵敏，可重现和准确的方法，其中qPCR引物与高度重复的宿主细胞基因组DNA序列互补 ，例如Alu等效序列。 本发明特别可用于确定要作为治疗剂例如治疗性蛋白质施用的生物制品中的残留基因组DNA的水平。
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