resumo
Green fluorescent protein (GFP) is a useful biomolecule in biotechnology; however, its price makes its widespread application prohibitive. To overcome this issue, recently, the use of aqueous two-phase systems (ATPS) for GFP purification was proposed as an alternative platform to reduce processing costs. Aligned with this goal, this study performed bioprocess modelling coupled with economic analysis using the software Biosolve to evaluate the potential and commercial applicability of ATPS for GFP purification. This work analysed a collection of fourteen ATPS to discriminate through production costs while also incorporating the concept of product purity into the calculations. The two best systems (a PEG-based and an ionic liquid (IL)-based ATPS) were placed in a full bioprocess at different scale models (1 to 100 L) to elucidate the viability of applying ATPS at large scale. Although the results showed that the PEG-based ATPS exhibit the lowest costs (between USD 3.5x10(3).g(-1) at 1 L and USD 0.33x10(3).g(-1) at 100 L), for further developments, the inclusion of an ATPS granting a higher purity is desired for the development of simpler bioprocesses. Therefore, as a third approach in this work, a sensitivity analysis was performed to determine the impact of varying different model parameters (recovery yield, material costs discount and production titre), to elucidate the circumstances under which the IL-based system can overcome the production costs of the traditional PEG-based ATPS. The results indicate that the best cost-effectiveness approach is to improve the production titre (although it can affect all ATPS studied), as an increase from 1.33 to 3.8 g/L is enough for the IL-based ATPS to be less expensive than the traditional system at all analysed scales. This study demonstrates that ATPS can greatly reduce GFP manufacturing costs, which can potentially help to popularize new applications of fluorescent proteins that are currently mostly restricted to research kits due to their high prices.
palavras-chave
AQUEOUS 2-PHASE EXTRACTION; B-PHYCOERYTHRIN; PURIFICATION; SEPARATION; SYSTEMS; GFP; UNCERTAINTY; STRATEGIES; INDUSTRY
categoria
Engineering, Chemical
autores
Torres-Acosta, MA; dos Santos, NV; Ventura, SPM; Coutinho, JAP; Rito-Palomares, M; Pereira, JFB
nossos autores
Jorge Fernando Brandão Pereira
Estudante de doutoramento
Sónia Patrícia Marques Ventura
Professor Auxiliaragradecimentos
This work was developed within the scope of the project Optimization and Scale-up of Novel Ionic-Liquid-based Purification Processes for Recombinant Green Fluorescent Protein produced by Escherichia coli, process 2014/19793-3, co-funded by FAPESP (Sao Paulo Research Foundation, Brazil) and FCT (Portuguese Foundation for Science and Technology, Portugal) reference of the project FAPESP/19793/2014 and of the project CICECO-Aveiro Institute of Materials, UIDB/50011/2020 & UIDP/50011/2020, financed by national funds through the Portuguese Foundation for Science and Technology/MCTES. This study was financed in part by the Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior -Brazil (CAPES) Finance Code 001. Nathalia V. Santos and J.F.B. Pereira acknowledge financial support from FAPESP through the projects 2016/07529-5 and 2014/16424-7, respectively.