@article{ ISI:000395956700030,
Author = {Silva, D. F. C. and Azevedo, A. M. and Fernandes, P. and Chu, V. and
   Conde, J. P. and Aires-Barros, M. R.},
Title = {{Determination of partition coefficients of biomolecules in a
   microfluidic aqueous two phase system platform using fluorescence
   microscopy}},
Journal = {{JOURNAL OF CHROMATOGRAPHY A}},
Year = {{2017}},
Volume = {{1487}},
Pages = {{242-247}},
Month = {{MAR 3}},
Abstract = {{Aqueous two phase systems (ATPS) offer great potential for selective
   separation of a wide range of biomolecules by exploring differences in
   molecular solubility in each of the two immiscible phases. However, ATPS
   use has been limited due to the difficulty in predicting the behavior of
   a given biomolecule in the partition environment together with the
   empirical and time-consuming techniques that are used for the
   determination of partition and extraction parameters. In this work, a
   fast and novel technique based on a microfluidic platform and using
   fluorescence microscopy was developed to determine the partition
   coefficients of biomolecules in different ATPS. This method consists of
   using a microfluidic device with a single microchannel and three inlets.
   In two of the inlets, solutions containing the ATPS forming components
   were loaded while the third inlet was fed with the FITC tagged
   biomolecule of interest prepared in milli-Q water. Using fluorescence
   microscopy, it was possible to follow the location of the FITC-tagged
   biomolecule and, by simply varying the pumping rates of the solutions,
   to quickly test a wide variety of ATPS compositions. The ATPS system is
   allowed 4 min for stabilization and fluorescence micrographs are used to
   determine the partition coefficient.The partition coefficients obtained
   were shown to be consistent with results from macroscale ATPS partition.
   This process allows for faster screening of partition coefficients using
   only a few microliters of material for each ATPS composition and is
   amenable to automation. The partitioning behavior of several
   biomolecules with molecular weights (MW) ranging from 5.8 to 150 kDa,
   and isoelectric points (pI) ranging from 4.7 to 6.4 was investigated, as
   well as the effect of the molecular weight of the polymer ATPS
   component. (C) 2016 Elsevier B.V. All rights reserved.}},
DOI = {{10.1016/j.chroma.2016.12.036}},
ISSN = {{0021-9673}},
EISSN = {{1873-3778}},
ResearcherID-Numbers = {{Conde, Joao Pedro/F-8533-2012
   Chu, Virginia/I-6048-2014
   Fernandes, Pedro/G-8180-2012
   Azevedo, Ana Margarida/A-2985-2011
   Aires-Barros, Maria Raquel/E-6021-2010}},
ORCID-Numbers = {{Conde, Joao Pedro/0000-0002-5677-3024
   Chu, Virginia/0000-0002-5306-4409
   Fernandes, Pedro/0000-0003-0271-7796
   Azevedo, Ana Margarida/0000-0001-5659-466X
   Aires-Barros, Maria Raquel/0000-0002-1813-4280}},
Unique-ID = {{ISI:000395956700030}},
}

{"_id":"AXD2GWw8jpMQSn3pv","bibbaseid":"silva-azevedo-fernandes-chu-conde-airesbarros-determinationofpartitioncoefficientsofbiomoleculesinamicrofluidicaqueoustwophasesystemplatformusingfluorescencemicroscopy-2017","authorIDs":[],"author_short":["Silva, D. F. C.","Azevedo, A. M.","Fernandes, P.","Chu, V.","Conde, J. P.","Aires-Barros, M. R."],"bibdata":{"bibtype":"article","type":"article","author":[{"propositions":[],"lastnames":["Silva"],"firstnames":["D.","F.","C."],"suffixes":[]},{"propositions":[],"lastnames":["Azevedo"],"firstnames":["A.","M."],"suffixes":[]},{"propositions":[],"lastnames":["Fernandes"],"firstnames":["P."],"suffixes":[]},{"propositions":[],"lastnames":["Chu"],"firstnames":["V."],"suffixes":[]},{"propositions":[],"lastnames":["Conde"],"firstnames":["J.","P."],"suffixes":[]},{"propositions":[],"lastnames":["Aires-Barros"],"firstnames":["M.","R."],"suffixes":[]}],"title":"Determination of partition coefficients of biomolecules in a microfluidic aqueous two phase system platform using fluorescence microscopy","journal":"JOURNAL OF CHROMATOGRAPHY A","year":"2017","volume":"1487","pages":"242-247","month":"MAR 3","abstract":"Aqueous two phase systems (ATPS) offer great potential for selective separation of a wide range of biomolecules by exploring differences in molecular solubility in each of the two immiscible phases. However, ATPS use has been limited due to the difficulty in predicting the behavior of a given biomolecule in the partition environment together with the empirical and time-consuming techniques that are used for the determination of partition and extraction parameters. In this work, a fast and novel technique based on a microfluidic platform and using fluorescence microscopy was developed to determine the partition coefficients of biomolecules in different ATPS. This method consists of using a microfluidic device with a single microchannel and three inlets. In two of the inlets, solutions containing the ATPS forming components were loaded while the third inlet was fed with the FITC tagged biomolecule of interest prepared in milli-Q water. Using fluorescence microscopy, it was possible to follow the location of the FITC-tagged biomolecule and, by simply varying the pumping rates of the solutions, to quickly test a wide variety of ATPS compositions. The ATPS system is allowed 4 min for stabilization and fluorescence micrographs are used to determine the partition coefficient.The partition coefficients obtained were shown to be consistent with results from macroscale ATPS partition. This process allows for faster screening of partition coefficients using only a few microliters of material for each ATPS composition and is amenable to automation. The partitioning behavior of several biomolecules with molecular weights (MW) ranging from 5.8 to 150 kDa, and isoelectric points (pI) ranging from 4.7 to 6.4 was investigated, as well as the effect of the molecular weight of the polymer ATPS component. (C) 2016 Elsevier B.V. All rights reserved.","doi":"10.1016/j.chroma.2016.12.036","issn":"0021-9673","eissn":"1873-3778","researcherid-numbers":"Conde, Joao Pedro/F-8533-2012 Chu, Virginia/I-6048-2014 Fernandes, Pedro/G-8180-2012 Azevedo, Ana Margarida/A-2985-2011 Aires-Barros, Maria Raquel/E-6021-2010","orcid-numbers":"Conde, Joao Pedro/0000-0002-5677-3024 Chu, Virginia/0000-0002-5306-4409 Fernandes, Pedro/0000-0003-0271-7796 Azevedo, Ana Margarida/0000-0001-5659-466X Aires-Barros, Maria Raquel/0000-0002-1813-4280","unique-id":"ISI:000395956700030","bibtex":"@article{ ISI:000395956700030,\nAuthor = {Silva, D. F. C. and Azevedo, A. M. and Fernandes, P. and Chu, V. and\n Conde, J. P. and Aires-Barros, M. R.},\nTitle = {{Determination of partition coefficients of biomolecules in a\n microfluidic aqueous two phase system platform using fluorescence\n microscopy}},\nJournal = {{JOURNAL OF CHROMATOGRAPHY A}},\nYear = {{2017}},\nVolume = {{1487}},\nPages = {{242-247}},\nMonth = {{MAR 3}},\nAbstract = {{Aqueous two phase systems (ATPS) offer great potential for selective\n separation of a wide range of biomolecules by exploring differences in\n molecular solubility in each of the two immiscible phases. However, ATPS\n use has been limited due to the difficulty in predicting the behavior of\n a given biomolecule in the partition environment together with the\n empirical and time-consuming techniques that are used for the\n determination of partition and extraction parameters. In this work, a\n fast and novel technique based on a microfluidic platform and using\n fluorescence microscopy was developed to determine the partition\n coefficients of biomolecules in different ATPS. This method consists of\n using a microfluidic device with a single microchannel and three inlets.\n In two of the inlets, solutions containing the ATPS forming components\n were loaded while the third inlet was fed with the FITC tagged\n biomolecule of interest prepared in milli-Q water. Using fluorescence\n microscopy, it was possible to follow the location of the FITC-tagged\n biomolecule and, by simply varying the pumping rates of the solutions,\n to quickly test a wide variety of ATPS compositions. The ATPS system is\n allowed 4 min for stabilization and fluorescence micrographs are used to\n determine the partition coefficient.The partition coefficients obtained\n were shown to be consistent with results from macroscale ATPS partition.\n This process allows for faster screening of partition coefficients using\n only a few microliters of material for each ATPS composition and is\n amenable to automation. The partitioning behavior of several\n biomolecules with molecular weights (MW) ranging from 5.8 to 150 kDa,\n and isoelectric points (pI) ranging from 4.7 to 6.4 was investigated, as\n well as the effect of the molecular weight of the polymer ATPS\n component. (C) 2016 Elsevier B.V. All rights reserved.}},\nDOI = {{10.1016/j.chroma.2016.12.036}},\nISSN = {{0021-9673}},\nEISSN = {{1873-3778}},\nResearcherID-Numbers = {{Conde, Joao Pedro/F-8533-2012\n Chu, Virginia/I-6048-2014\n Fernandes, Pedro/G-8180-2012\n Azevedo, Ana Margarida/A-2985-2011\n Aires-Barros, Maria Raquel/E-6021-2010}},\nORCID-Numbers = {{Conde, Joao Pedro/0000-0002-5677-3024\n Chu, Virginia/0000-0002-5306-4409\n Fernandes, Pedro/0000-0003-0271-7796\n Azevedo, Ana Margarida/0000-0001-5659-466X\n Aires-Barros, Maria Raquel/0000-0002-1813-4280}},\nUnique-ID = {{ISI:000395956700030}},\n}\n\n","author_short":["Silva, D. F. C.","Azevedo, A. M.","Fernandes, P.","Chu, V.","Conde, J. P.","Aires-Barros, M. R."],"key":"ISI:000395956700030","id":"ISI:000395956700030","bibbaseid":"silva-azevedo-fernandes-chu-conde-airesbarros-determinationofpartitioncoefficientsofbiomoleculesinamicrofluidicaqueoustwophasesystemplatformusingfluorescencemicroscopy-2017","role":"author","urls":{},"downloads":0},"bibtype":"article","biburl":"http://nanotec.cnr.it/data/nanotec/nanotec-full.bib","creationDate":"2020-04-21T16:04:30.784Z","downloads":0,"keywords":[],"search_terms":["determination","partition","coefficients","biomolecules","microfluidic","aqueous","two","phase","system","platform","using","fluorescence","microscopy","silva","azevedo","fernandes","chu","conde","aires-barros"],"title":"Determination of partition coefficients of biomolecules in a microfluidic aqueous two phase system platform using fluorescence microscopy","year":2017,"dataSources":["ZTEbWc6bW5f9DTsjF"]}