The localization of replication origins on ARS plasmids in S. Individual-specific 'fingerprints' of human DNA. The rabbit β-globin gene contains a large large insert in the coding sequence. Polymorphism of DNA sequence adjacent to human beta-globin structural gene: relationship to sickle mutation. Construction of a genetic linkage map in man using restriction fragment length polymorphisms. Botstein, D., White, R.L., Skolnick, M.The arrangement of simian virus 40 sequences in the DNA of transformed cells. A membrane-filter technique for the detection of complementary DNA. Detection of specific sequences among DNA fragments separated by gel electrophoresis. Method for detection of specific RNAs in agarose gels by transfer to diazobenzyloxymethyl-paper and hybridization with DNA probes. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. An improved procedure for starch-gel electrophoresis: further variations in the serum proteins of normal individuals. The determination of the molecular weight of ribonucleic acid by polyacrylamide-gel electrophresis. Specific cleavage of simian virus 40 DNA by restriction endonuclease of Hemophilus influenzae. Sartorius uses the ABI 3500 platform for traditional Sanger sequencing building on more than 25 years’ experience in nucleic acid sequencing with a wide array of different organisms and sample types including recombinant proteins from both human and CHO cells. Sequencing of the flanking region (at the DNA level) is recommended by ICH guidelines and should be considered prior to phase 3 clinical trials. In addition to the coding region, the flanking region of the transgene is also an important consideration as this can affect the productivity and expression levels. This reveals evidence confirming there are no changes to the genetic code and the cells have not undergone any significant changes. Sartorius offers the following genetic stability assays and can advise on the most appropriate approach for your product:Ĭell Line Identity Testing (DNA Barcoding, RAPD)ĭetermining the nucleic acid sequence of the transgene used to produce the protein of interest is one of the critical tests carried out to determine the genetic stability of the cell line.ĭuring early stages of the development of a recombinant protein drug, the messenger RNA (mRNA) sequence of the transgene is determined for both the Master Cell Bank (MCB) and End of Production Cell (EOP) Bank. These tests take place during the characterization of a production cell line, and to assess the genetic stability of the Master Cell Bank and the End of Production Cell Bank. Testing can also be carried out at other times, but is particularly useful during phases where there are changes to the fermentation process. There are multiple techniques to determine the genetic stability of a cell line. ICH Guidelines state the stability and integrity of the insertion sequence needs to be characterized to ensure stable production of the correct protein. This is just one of the required steps ensure the cell line and production system is acceptable. Genetic stability is a specific characterization of the cells used in the production of a biologic. Genetically modified cells contain transgene insertion sequences that code for the protein of interest. Transgenes have the potential to mutate, which can impact the quality of the product or the proper execution of processes. Genetic Stability Analysis: Characterization and Stability Testing
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