Soares R.R.G.,University of Lisbon |
Silva D.F.C.,University of Lisbon |
Fernandes P.,University of Lisbon |
Azevedo A.M.,University of Lisbon |
And 3 more authors.
Biotechnology Journal | Year: 2016
Aqueous two-phase extraction (ATPE) is a biocompatible liquid-liquid (L-L) separation technique that has been under research for several decades towards the purification of biomolecules, ranging from small metabolites to large animal cells. More recently, with the emergence of rapid-prototyping techniques for fabrication of microfluidic structures with intricate designs, ATPE gained an expanded range of applications utilizing physical phenomena occurring exclusively at the microscale. Today, research is being carried simultaneously in two different volume ranges, mL-scale (microtubes) and nL-scale (microchannels). The objective of this review is to give insight into the state of the art at both microtube and microchannel-scale and to analyze whether miniaturization is currently a competing or divergent technology in a field of applications including bioseparation, bioanalytics, enhanced fermentation processes, catalysis, high-throughput screening and physical/chemical compartmentalization. From our perspective, both approaches are worthy of investigation and, depending on the application, it is likely that either (i) one of the approaches will eventually become obsolete in particular research areas such as purification at the preparative scale or high-throughput screening applications; or (ii) both approaches will function as complementing techniques within the bioanalytics field. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim..
Bras E.J.,University of Lisbon |
Chu V.,Institute Engineering Of Sistemas E Computadores Microsistemas E Nanotecnologias Inesc Mn And In Institute Of Nanoscience And Nanotechnology Lisbon Portugal |
Aires-Barros M.R.,University of Lisbon |
Conde J.P.,University of Lisbon |
Fernandes P.,University of Lisbon
Journal of Chemical Technology and Biotechnology | Year: 2016
BACKGROUND: The rapidly increasing use of biotechnology in many industries has led to the need for novel methods for cell culture which provide an efficient way to either optimize or perform fermentation operations. In parallel, microfabrication techniques allowed the development of microfluidic chips for complex handling of fluids and cells. RESULTS: This work presents a microfluidic platform to trap non-adherent cells for the continuous production of a biomolecule of interest. The biological system chosen as a model was the yeast species Saccharomyces cerevisiae and the extracellular protein invertase. The use of the appropriate combination of the flow rate of the medium, medium dilution rate, and pH allowed effective control of the cell growth in the microfluidic bioreactor while at the same time maximizing the invertase activity per cell. The microfluidic bioreactor allowed for continuous cell culture for 32 h and its productivity both per cell (3.22 × 10-8 U cell-1) and per consumed nutrient (3.79 U mg-1 sucrose) was consistently higher than its macroscale batch and continuous reactor counterparts. CONCLUSION: This work demonstrated that a microfluidic bioreactor can be used for continuous production of an extracellular protein using hydrodynamically trapped non-adherent yeast cells. © 2016 Society of Chemical Industry.