Title
Miniaturization of aqueous two-phase extraction for biological applications
Conference Dates
March 5-10, 2017
Abstract
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.
Studies of ATPSs at nanoliter-scale are further extending the range of applications of these systems by taking advantage of rapid diffusion times, increased degree of control of individual liquid streams and droplets, continuous flow and the integration of multi-dimensional separation modes. Several examples of microfluidic ATPS platforms are described.
The partition of molecules between two co-flowing liquid streams confined within a microchannel was successfully demonstrated by the on-line extraction of a fluorescein isothiocyanate (FITC) labeled immunoglobulin G (IgG) from a salt rich flow to a PEG rich flow. IgG diffusion to the PEG-rich phase was complete after 16 cm of channel using flow rates of 1 and 0.2 μL/min for the salt and PEGrich phases respectively. Besides proteins, ATPS have also been used to separate other more complex biomolecules in microfluidics such as virus-like particles.
The potential of miniaturization as a high-throughput screening tool has also been explored. The developed setup allowed the screening of a wide range of concentrations inside the microchannel by varying the flow rates of the solutions while using sub-mL volumes for each ATPS-forming system.
As a novel demonstration of the integrative potential of ATPE as a microfluidic sample preparation module, a microfluidic device comprising two modules was developed and used to perform a complex matrix clean-up in-line with an immunoassay.
References:
Silva, D. F., Azevedo, A. M., Fernandes, P., Chu, V. et al., Design of a microfluidic platform for monoclonal antibody extraction using an aqueous two-phase system. J. Chromatogr. A 2012, 1249, 1–7.
Jacinto, M. J., Soares, R. R. G., Azevedo, A. M., Chu, V. et al., Optimization and miniaturization of aqueous two phase systems for the purification of recombinant human immunodeficiency virus-like particles from a CHO cell supernatant. Sep. Purif. Technol. 2015, 154, 27–35.
Silva, D. F. C., Azevedo, A. M., Fernandes, P., Chu, V. et al., Determination of aqueous two phase system binodal curves using a microfluidic device. J. Chromatogr. A 2014, 1370, 115–120.
Soares, R. R., Novo, P., Azevedo, A. M., Fernandes, P. et al., On-chip sample preparation and analyte quantification using a microfluidic aqueous two-phase extraction coupled with an immunoassay. Lab Chip 2014, 14, 4284–4294
Recommended Citation
Raquel Aires-Barros, Ruben R.G. Soares, Maria Joao Jacinto, Daniel Silva, Virginia Chu, Ana M. Azevedo, and João P. Conde, "Miniaturization of aqueous two-phase extraction for biological applications" in "Separations Technology IX: New Frontiers in Media, Techniques, and Technologies", Kamalesh K. Sirkar, New Jersey Institute of Technology, USA Steven M. Crame, Rensselaer Polytechnic Institute, USA João G. Crespo, LAQV-Requimte, FCT-Universidade Nova de Lisboa, Caparica, Portugal Marco Mazzotti, ETH Zurich, Switzerland Eds, ECI Symposium Series, (2017). https://dc.engconfintl.org/separations_technology_ix/36