Plasmid DNA for biopharmaceutical applications is produced easily in Escherichia coli bacteria. The cell lysis is the most crucial step for purification of plasmid DNA. In this paper, we describe a continuous cell alkaline lysis, neutralization, and clarification combination process for production o
Time Course of SDS–alkaline lysis of recombinant bacterial cells for plasmid release
✍ Scribed by L. A. S. Ciccolini; P. Ayazi Shamlou; N. J. Titchener-Hooker; J. M. Ward; P. Dunnill
- Publisher
- John Wiley and Sons
- Year
- 1998
- Tongue
- English
- Weight
- 89 KB
- Volume
- 60
- Category
- Article
- ISSN
- 0006-3592
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✦ Synopsis
SDS-alkaline lysis of recombinant Escherichia coli cell suspensions was carried out in a coaxial cylinder rheometer, and the data were used to establish the time course of lysis reaction. The results of the experiments showed that cell lysis reaction time depended on cell strain but was unaffected by plasmid size and plasmid copy number. The high molecular weight globular proteins and chromosomal DNA were denatured, and the resulting changes in rheometric measurements characterised the denaturation time.
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## Abstract A state‐of‐the‐art in‐line static mixer reactor (ISMR) was invented to lyse __E. coli__ cells and neutralize the cell lysate continuously and efficiently for the extraction of plasmid DNA. It comprised two connected static dynamic mixers, each 0.01 m in diameter and 0.9 m in length, one