Browsing by Author "Gomes, Maria João Teles"
Now showing 1 - 1 of 1
Results Per Page
Sort Options
- Nanodiamonds selectivity towards DNA and RNA: exploiting a tool for biopharmaceuticals capturePublication . Gomes, Maria João Teles; Sousa, Fani Pereira de; Silva, Láudia Sofia Castro Gomes da; Ferreira, Pedro Filipe LopesThe therapeutic potential of RNA has been unveiled since its discovery in the beginning of the 20th century. Notwithstanding, for RNA to be used for such applications, it has to present high purity, integrity, and biological activity. The recombinant production of nucleic acids is a complex process involving multiple steps. It can be divided into upstream and downstream processing. While improvements have been made in upstream processing, the same level of progress is not directly observed for downstream processing. Downstream processing, a crucial part of a bioprocess, involves the recovery, concentration, and purification of the product from the upstream process. It encompasses three main steps: initial recovery, high-resolution purification, and polishing. The higher the product purity level achieved in the low-resolution purification phase, the greater the potential for reducing high-resolution purification steps, thereby enabling cost reduction. This is particularly important, as downstream processing accounts for nearly 80% of the total biopharmaceutical production costs. To advance on the current state-of-art for RNA purification, an optimization of an adsorption method for the pre-purification of RNA when it is present in complex E. coli lysate samples, using MDs, NDs, and functionalized NDs as adsorbents was carried out. A simple and efficient method, using the CM that showed to be the most promising, for the selective capture and recovery of RNA from E. coli lysates containing impurities such as pDNA and proteins, was developed. Particularly, ND-ox were the materials that showed to be the most promising, allowing the achievement of an adsorption capacity of 86.9 mg of RNA/g of CM, not being present pDNA in the RNA sample recovered at the end of the procedure, and achieving an elimination of solubilized proteins of 91.28 % relatively to the initial sample. Globally, the results of this work demonstrated that ND-ox can be used as adsorbents in DSPE, being capable of selectively capturing RNA and enabling its complete recovery without contamination.