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:
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:
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.