método de purificação de plasmìdeo dna

• Main Authors: MICHEL COUDERT, FRANCIS BLANCHE, NICOLAS MAESTRALI, THIERRY GUILLEMIN, DAVID GAILLAC
• Format: Patent
• Language: Portuguese
• Published: 11.09.2007
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; COMPOSITIONS THEREOF; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; CULTURE MEDIA; ENZYMOLOGY; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; PROCESSES OF PREPARING SUCH COMPOSITIONS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

MéTODO DE PURIFICAçãO DE PLASMìDEO DNA. Esta invenção proporciona um processo para a lisis alcalina contínua de uma suspensão bacterial de modo a produzir pDNA. Proporciona-se adicionalmente etapas de purificação opcionais adicionais, incluindo filtração de lisato, cromatografia de troca de ânion, cromatografia de afinidade de triplex e cromatografia de interação hidrofóbica. Estas etapas de purificação opcionais podem ser combinadas com a lisis contínua de modo a produzir um produto de pDNA altamente purificado substancialmente livre de gDNA, RNA, proteína, endotoxin e outros contaminantes. This invention provides a process for the continuous alkaline lysis of a bacterial suspension in order to harvest pDNA. It further provides for optional additional purification steps, including lysate filtration, anion exchange chromatography, triplex affinity chromatogragphy, and hydrophobic interaction chromatography. These optional purification steps can be combined with the continuous lysis in order to produce a highly purified pDNA product substantially free of gDNA, RNA, protein, endotoxin, and other contaminants.