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6 results on '"Aqueous two-phase micellar systems (ATPMS)"'

1. Effect of electrolytes as adjuvants in GFP and LPS partitioning on aqueous two-phase systems: 2. Nonionic micellar systems.

Author

Teixeira-Pinto, Renata Garcia Rodrigues, Molino, João Vitor Dutra, Santos-Ebinuma, Valéria Carvalho, Pessoa, Adalberto, Valentini, Sandro Roberto, Pereira, Jorge Fernando Brandão, and Lopes, André Moreni

Subjects
*MICELLES, *BIOPHARMACEUTICS, *GREEN fluorescent protein
Abstract

Graphical abstract Highlights • GFP recovery and LPS removal on C 10 E 4 /buffer + salts-based ATPMS were studied. • C 10 E 4 /buffer + salts promoted GFP partitioning to the micelle-poor phase (K GFP > 7). • C 10 E 4 /buffer + salts-based ATPMS removed LPS to the micelle-rich phase (REM LPS > 98%). • High loads of LPS (>106 EU/mL) increased the GFP partition into micelle-rich phase. • C 10 E 4 /buffer + salt are able to separate selectively the GFP from contaminant LPS. Abstract Lipopolysaccharide endotoxins (LPS) are the most common contaminant pyrogenic compounds found in intracellular recombinant biomolecules purified from Gram-negative bacteria, such as Escherichia coli. Thus, the purification downstream processing should guarantee the effective removal of LPS from the final bioproduct, particularly, therapeutic biopharmaceuticals. Aqueous two-phase micellar systems (ATPMS) appear to be an excellent strategy to purify recombinant biopharmaceuticals from the cell lysate of E. coli , reducing high LPS concentrations. In order to demonstrate the effectiveness of ATPMS as a biopharmaceutical purification platform, the influence of inorganic salt electrolytes (NaCl, Li 2 SO 4 , KI, or KNO 3) on the partitioning of green fluorescent protein (GFP) and LPS removal using ATPMS composed of n -decyl tetraethylene oxide (C 10 E 4) was evaluated. The impact of different LPS concentrations on GFP partitioning was also studied. The addition of electrolytes (i.e., NaCl or Li 2 SO 4) to the C 10 E 4 -based ATPMS have reduced the phase forming temperatures to very mild conditions (ca. 17.00 and 13.00 °C, for NaCl and Li 2 SO 4 , respectively). The selective partitioning ability of the proposed ATPMS was further demonstrated, where a complete removal of the LPS from the micelle-poor phase (REM LPS > 98%) and a preferential GFP recovery (REC GFP = 97%, K GFP > 7) to the micelle-poor phase was obtained. The GFP partitioning was even enhanced by increasing LPS loading (104–106 EU/mL), probably due to the formation of mixed micelles between LPS and C 10 E 4. It is here demonstrated that a C 10 E 4 /buffer + salt-based ATPMS can be a useful and straightforward platform for the removal of endotoxin contaminants and the purification of recombinant biopharmaceuticals from E. coli cell lysates. [ABSTRACT FROM AUTHOR]

Published
2019
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2. Amphiphilic copolymer aqueous solutions with cholinium ionic liquids as adjuvants: New insights into determination of binodal curves and phase-separation mechanisms

Author

Jorge F. B. Pereira, Cassamo Ussemane Mussagy, Isabelle S. Kurnik, André Moreni Lopes, Universidade Estadual Paulista (Unesp), CIEPQPF, and Universidade Estadual de Campinas (UNICAMP)

Subjects
Materials science, Phase separation, 02 engineering and technology, 010402 general chemistry, 01 natural sciences, Lower critical solution temperature, chemistry.chemical_compound, Temperature modulated optical refractometry (TMOR), Pulmonary surfactant, Ionic liquids (ILs), Materials Chemistry, Physical and Theoretical Chemistry, Aqueous two-phase micellar systems (ATPMS), Spectroscopy, Binodal, Cloud-point temperature, Aqueous solution, 021001 nanoscience & nanotechnology, Condensed Matter Physics, Atomic and Molecular Physics, and Optics, Phase formation, 0104 chemical sciences, Electronic, Optical and Magnetic Materials, Refractometry, chemistry, Chemical engineering, Ionic liquid, 0210 nano-technology, Amphiphilic copolymer
Abstract

Made available in DSpace on 2021-06-25T10:11:10Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-11-15 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Aqueous two-phase micellar systems (ATPMS) are more than simple amphiphilic copolymer or surfactant aqueous solutions since due to the formation of biphasic regimes, at certain temperatures, these can be successfully used for the separation, encapsulation, and formulation of several bio-based compounds. The recent use of ionic liquids (ILs) as adjuvants in the formation of ATPMS have enlarged its “working window”, but the measurement of respective binodal curves is still very empiric and dependent of operator accuracy through the visual determination of cloud-point temperature. In this work, the range of ATPMS using ILs as adjuvants was extended, providing more insights about their phase formation mechanisms, as well as into determination of cloud-point values of the lower critical solution temperature (LCST)-type demixing transition by using the new temperature modulated optical refractometry approach. The binodal curves of ATPMS composed of Pluronic® L35/PBS buffer at pH 7.4 + cholinium ILs were determined, by using two different methods, i.e.: cloud-point temperature (TCP) visual determination versus temperature modulated optical refractometry (TMOR). TMOR method was more sensitive to the determination of biphasic transition, particularly, in highly diluted solutions (< 3.0 wt% of Pluronic L35). The results also established that the addition of cholinium ILs decreased significantly the phase separation temperatures of Pluronic L35/PBS, with an increase of biphasic region (viz. lower of TCP values) as a function of the relative hydrophobicity of the IL anion, i.e.: [Hex]− > [But]− > [Pro]− > [Ac]− > Cl−. The use of cholinium ILs as adjuvants is particularly important to provide mild conditions for ATPMS-based separation and encapsulation strategies, while TMOR-based methods allow an improvement of phase separation determination accuracy. Department of Engineering of Bioprocesses and Biotechnology School of Pharmaceutical Sciences São Paulo State University (UNESP) University of Coimbra CIEPQPF Department of Chemical Engineering, Rua Sílvio Lima, Pólo II – Pinhal de Marrocos Faculty of Pharmaceutical Sciences University of Campinas Department of Engineering of Bioprocesses and Biotechnology School of Pharmaceutical Sciences São Paulo State University (UNESP) CAPES: 001 FAPESP: 2014/16424-7 FAPESP: 2017/10789-1 FAPESP: 2018/05111-9 FAPESP: 2018/10799-0 FAPESP: 2019/05624-9 FAPESP: 2019/08549-8

Published
2020

3. Separation and purification of curcumin using novel aqueous two-phase micellar systems composed of amphiphilic copolymer and cholinium ionic liquids

Author

Mariana A. Noronha, Priscila Gava Mazzola, André Moreni Lopes, António A. Vicente, Jorge F. B. Pereira, Mayra C.C. Câmara, Isabelle S. Kurnik, Universidade Estadual Paulista (Unesp), Universidade Estadual de Campinas (UNICAMP), University of Minho, Rua Sílvio Lima, and Universidade do Minho

Subjects
separation, Filtration and Separation, 02 engineering and technology, Analytical Chemistry, Ion, chemistry.chemical_compound, 020401 chemical engineering, Phase (matter), Copolymer, 0204 chemical engineering, chemistry.chemical_classification, Aqueous solution, Science & Technology, ionic liquids (ILs), Biomolecule, aqueous two-phase micellar systems (ATPMS), Poloxamer, 021001 nanoscience & nanotechnology, chemistry, Ionic liquid, Pluronic F68, polymeric micelles (PMs), 0210 nano-technology, Selectivity, curcumin (CCM), Nuclear chemistry
Abstract

Novel aqueous two-phase micellar systems (ATPMS) composed of Pluronic F68, a triblock amphiphilic copolymer, and cholinium-based ionic liquids (ILs) were formulated and applied for separation/purification of curcumin (CCM). CCM stability in the presence of ATPMS components was also evaluated. CCM is stable up to 24 h in copolymer (1.0 10.0 wt%) and ILs (0.1 3.0 M) aqueous solutions. Very mild phase separation conditions (close to room temperature) were achieved by adding cholinium ILs to the Pluronic F68 + McIlvaine buffer at pH 6.0 solution. The decrease of cloud-point temperature is dependent on the relative hydrophobicity of IL anion, [Hex] > [But] > [Prop] > [Ac] > Cl. ATPMS composed of more hydrophobic ILs ([Ch][Hex] > [Ch][But] > [Ch][Prop]) are most efficient in the partition of commercial CCM into polymeric micelles-rich phase. The best ATPMS (0.70 M [Ch][But] and 0.60 M [Ch][Hex]-based ATPMS) were then used to purify CCM from a crude extract of Curcuma longa L. Both systems were very selective to separate CCM from protein-based contaminants (selectivity values 25; purification yields 12-fold). Pluronic F68-based ATPMS are promising for selective separation of hydrophobic biomolecules by using cholinium-based ILs as adjuvants to adjust phase separation temperatures and biomolecules partition., This study was funded by the Coordination for Higher Level Graduate Improvements (CAPES/Brazil, finance code 001), National Council for Scientific and Technological Development (CNPq/Brazil) and the State of São Paulo Research Foundation (FAPESP/Brazil, processes #2014/16424-7, #2017/10789-1, #2018/10799-0, #2018/05111-9; #2019/05624-9, and #2019/08549-8). A.M. Lopes and J.F.B. Pereira are grateful for the language revision of native speaker H.S. Pacheco Neto., info:eu-repo/semantics/publishedVersion

Published
2020

4. Effect of electrolytes as adjuvants in GFP and LPS partitioning on aqueous two-phase systems: 2. Nonionic micellar systems

Author

André Moreni Lopes, Jorge F. B. Pereira, Adalberto Pessoa, Renata Garcia Rodrigues Teixeira-Pinto, Valéria de Carvalho Santos-Ebinuma, João Vitor Dutra Molino, Sandro Roberto Valentini, Universidade de São Paulo (USP), Ronin Institute, and Universidade Estadual Paulista (Unesp)

Subjects
Lysis, Filtration and Separation, 02 engineering and technology, medicine.disease_cause, Micelle, Lipopolysaccharide (LPS) removal, Analytical Chemistry, law.invention, Green fluorescent protein, 020401 chemical engineering, law, PROTEÍNAS DE FLUORESCÊNCIA VERDE, medicine, 0204 chemical engineering, Aqueous two-phase micellar systems (ATPMS), Escherichia coli, Inorganic salts, Aqueous solution, Chromatography, Downstream processing, biology, Chemistry, 021001 nanoscience & nanotechnology, biology.organism_classification, Biopharmaceutical purification, Green fluorescent protein (GFP), Recombinant DNA, 0210 nano-technology, C10E4, Bacteria
Abstract

Made available in DSpace on 2019-10-06T15:18:27Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-02-08 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Lipopolysaccharide endotoxins (LPS) are the most common contaminant pyrogenic compounds found in intracellular recombinant biomolecules purified from Gram-negative bacteria, such as Escherichia coli. Thus, the purification downstream processing should guarantee the effective removal of LPS from the final bioproduct, particularly, therapeutic biopharmaceuticals. Aqueous two-phase micellar systems (ATPMS) appear to be an excellent strategy to purify recombinant biopharmaceuticals from the cell lysate of E. coli, reducing high LPS concentrations. In order to demonstrate the effectiveness of ATPMS as a biopharmaceutical purification platform, the influence of inorganic salt electrolytes (NaCl, Li2SO4, KI, or KNO3) on the partitioning of green fluorescent protein (GFP) and LPS removal using ATPMS composed of n-decyl tetraethylene oxide (C10E4) was evaluated. The impact of different LPS concentrations on GFP partitioning was also studied. The addition of electrolytes (i.e., NaCl or Li2SO4) to the C10E4-based ATPMS have reduced the phase forming temperatures to very mild conditions (ca. 17.00 and 13.00 °C, for NaCl and Li2SO4, respectively). The selective partitioning ability of the proposed ATPMS was further demonstrated, where a complete removal of the LPS from the micelle-poor phase (REMLPS > 98%) and a preferential GFP recovery (RECGFP = 97%, KGFP > 7) to the micelle-poor phase was obtained. The GFP partitioning was even enhanced by increasing LPS loading (104–106 EU/mL), probably due to the formation of mixed micelles between LPS and C10E4. It is here demonstrated that a C10E4/buffer + salt-based ATPMS can be a useful and straightforward platform for the removal of endotoxin contaminants and the purification of recombinant biopharmaceuticals from E. coli cell lysates. Department of Biochemical and Pharmaceutical Technology School of Pharmaceutical Sciences University of São Paulo – FCF/USP Ronin Institute Department of Bioprocesses and Biotechnology School of Pharmaceutical Sciences São Paulo State University – UNESP Department of Bioprocesses and Biotechnology School of Pharmaceutical Sciences São Paulo State University – UNESP FAPESP: #2005/60159-7 FAPESP: #2007/51978-0 FAPESP: #2014/19793-3 CAPES: 0366/09-9

Published
2019

5. Amphiphilic copolymer aqueous solutions with cholinium ionic liquids as adjuvants: New insights into determination of binodal curves and phase-separation mechanisms.

Author

Kurnik, Isabelle S., Mussagy, Cassamo U., Pereira, Jorge F.B., and Lopes, André M.

Subjects
*IONIC solutions, *AQUEOUS solutions, *BIOSURFACTANTS, *PHASE separation, *CRITICAL temperature, *CURVES, *POLYBUTENES, *IONIC liquids
Abstract

Aqueous two-phase micellar systems (ATPMS) are more than simple amphiphilic copolymer or surfactant aqueous solutions since due to the formation of biphasic regimes, at certain temperatures, these can be successfully used for the separation, encapsulation, and formulation of several bio-based compounds. The recent use of ionic liquids (ILs) as adjuvants in the formation of ATPMS have enlarged its "working window", but the measurement of respective binodal curves is still very empiric and dependent of operator accuracy through the visual determination of cloud-point temperature. In this work, the range of ATPMS using ILs as adjuvants was extended, providing more insights about their phase formation mechanisms, as well as into determination of cloud-point values of the lower critical solution temperature (LCST)-type demixing transition by using the new temperature modulated optical refractometry approach. The binodal curves of ATPMS composed of Pluronic® L35/PBS buffer at pH 7.4 + cholinium ILs were determined, by using two different methods, i.e.: cloud-point temperature (T CP) visual determination versus temperature modulated optical refractometry (TMOR). TMOR method was more sensitive to the determination of biphasic transition, particularly, in highly diluted solutions (< 3.0 wt% of Pluronic L35). The results also established that the addition of cholinium ILs decreased significantly the phase separation temperatures of Pluronic L35/PBS, with an increase of biphasic region (viz. lower of T CP values) as a function of the relative hydrophobicity of the IL anion, i.e. : [Hex]− > [But]− > [Pro]− > [Ac]− > Cl−. The use of cholinium ILs as adjuvants is particularly important to provide mild conditions for ATPMS-based separation and encapsulation strategies, while TMOR-based methods allow an improvement of phase separation determination accuracy. Unlabelled Image • Refractometry index Vs. cloud-point temperature by visual technique were compared. • The refractometry-based technique is more accurate to determine the binodal curves. • Cholinium ILs decreased the phase separation temperature of Pluronic L35/PBS. • Cloud-point temperature decreased with the increase of the IL anion hydrophobicity. [ABSTRACT FROM AUTHOR]

Published
2020
Full Text
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6. Separation and purification of curcumin using novel aqueous two-phase micellar systems composed of amphiphilic copolymer and cholinium ionic liquids.

Author

Kurnik, Isabelle S., Noronha, Mariana A., Câmara, Mayra C.C., Mazzola, Priscila G., Vicente, António A., Pereira, Jorge F.B., and Lopes, André M.

Subjects
*IONIC liquids, *PHASE separation, *CHEMICAL purification, *TURMERIC, *AQUEOUS solutions, *MICELLAR solutions, *BIOMOLECULES, *MICELLES
Abstract

• Partition of curcumin (CCM) using ATPMS composed of Pluronic F68 + ILs was studied. • Cholinium ILs decreased the phase separation temperature of Pluronic F68/buffer. • The phase separation decreased with the increase of the IL anion hydrophobicity. • CCM was preferentially partitioned into polymeric micelles (PMs)-rich phase. • ILs-based ATPMS were very selective to recover CCM from crude extract of C. longa L. Novel aqueous two-phase micellar systems (ATPMS) composed of Pluronic F68, a triblock amphiphilic copolymer, and cholinium-based ionic liquids (ILs) were formulated and applied for separation/purification of curcumin (CCM). CCM stability in the presence of ATPMS components was also evaluated. CCM is stable up to 24 h in copolymer (1.0 – 10.0 wt%) and ILs (0.1 – 3.0 M) aqueous solutions. Very mild phase separation conditions (close to room temperature) were achieved by adding cholinium ILs to the Pluronic F68 + McIlvaine buffer at pH 6.0 solution. The decrease of cloud-point temperature is dependent on the relative hydrophobicity of IL anion, [Hex]− > [But]− > [Pro]− > [Ac]− > Cl−. ATPMS composed of more hydrophobic ILs ([Ch][Hex] > [Ch][But] > [Ch][Pro]) are most efficient in the partition of commercial CCM into polymeric micelles (PMs)-rich phase. The best ATPMS (0.70 M [Ch][But] and 0.60 M [Ch][Hex]-based ATPMS) were then used to purify CCM from a crude extract of Curcuma longa L. Both systems were very selective to separate CCM from protein-based contaminants (selectivity values ≥ 25; purification yields ≥ 12-fold). Pluronic F68-based ATPMS are promising for selective separation of hydrophobic biomolecules by using cholinium-based ILs as adjuvants to adjust phase separation temperatures and biomolecules' partition. [ABSTRACT FROM AUTHOR]

Published
2020
Full Text
View/download PDF

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