Enhanced performance of polymer-polymer aqueous two-phase systems using ionic liquids as adjuvants towards the purification of recombinant proteins

Castro, Leonor S.; Pereira, Patrícia; Passarinha, Luís A.; Freire, Mara G.; Pedro, Ana

doi:10.1016/j.seppur.2020.117051

Cited by 42 publications

(31 citation statements)

References 61 publications

(83 reference statements)

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“…In this regard, the emergence of biopharmaceuticals has allowed tremendous improvements in life quality [ 2 ], being at the cornerstone of the progress achieved in the last decades on the prevention and treatment of a wide range of diseases (e.g., cancer, infectious diseases, neurodegenerative diseases, among others). Biopharmaceuticals, also called biotherapeutics or biologicals, are products of biological origin such as proteins, nucleic acids, blood-derived products, somatic cells, or derivatives that are produced or extracted from living sources (e.g., microorganisms, cells, plants, or animals) [ 3 , 4 , 5 ]. Nowadays, recombinant therapeutic proteins and antibodies are considered the most abundant types of biopharmaceutical products in the market [ 3 ].…”

Section: Clinical Importance Of Interferon-based Biopharmaceuticals and Market Overviewmentioning

confidence: 99%

“…Over the years, alternatives to the widely applied and effective column liquid chromatographic processes have been described, mostly aiming to overcome their high cost and limited capacity [ 85 ]. Consequently, new techniques have been suggested, such as aqueous two-phase systems (ATPS) [ 5 , 105 , 106 ], cationic surfactant-based reverse micellar extraction [ 107 ], and immunomagnetic microspheres [ 108 ], schematized in Figure 8 . These techniques aim for a high recovery yield and purity through the least possible number of steps, easy application on an industrial scale, a process that is human- and environmentally friendly, and a system that is cost-effective [ 4 , 61 , 84 ].…”

Section: Therapeutic Cloned Interferonsmentioning

confidence: 99%

“…It was observed that IFNα-2b tends to migrate to the PEG-rich phase (being the phase also enriched in IL), whereas the remaining proteins tend to precipitate at the interface (fitting within the three-phase partitioning approach). In comparison with the ATPS without adjuvant, most systems comprising ILs as adjuvants lead to enhancements in the purification factors of IFNα-2b—namely, from 2.28 up to 6.77—with extraction efficiencies above 90% [ 5 ]. The purity of IFNα-2b was found to be maximized using ILs composed of aromatic cations and anions with high hydrogen-bond basicity (1-butyl-3-methylimidazolium acetate, [C 4 mim][CH 3 COO]), and the secondary structure of the target protein was found to be preserved during the purification step.…”

Section: Therapeutic Cloned Interferonsmentioning

confidence: 99%

“…The purity of IFNα-2b was found to be maximized using ILs composed of aromatic cations and anions with high hydrogen-bond basicity (1-butyl-3-methylimidazolium acetate, [C 4 mim][CH 3 COO]), and the secondary structure of the target protein was found to be preserved during the purification step. Overall, this study demonstrated the ability of ILs to tune the characteristics of the ATPS coexisting phases toward improved purification processes by taking advantage of the designer solvent ability of ILs [ 5 ]. In summary, the described results indicate that optimized alternative purification platforms such as ATPS represent a promising technique for the recovery and purification of IFN.…”

Section: Therapeutic Cloned Interferonsmentioning

confidence: 99%

“…In cases where IFN is transported to the periplasm, selective disruption of the outer membrane is crucial to avoid complete lysis, thus ensuring that the target protein is recovered After cell lysis, inclusion bodies are isolated from soluble proteins by centrifugation [60] and are then subjected to different washing steps, e.g., with detergents (Tween 20 [89] or triton X-100 [60]), low concentrations of urea [60,90], or sodium deoxycholate [91]. Subsequently, solubilization occurs in a high concentration of denaturing agents such as urea (6-8 M) [92] or guanidine hydrochloride (6 M) [60,90] and can be enhanced at alkaline pH [5,49]. Milder solubilization methods employing 1-propanol or 2-butanol in the presence of sodium dodecyl sulfate (SDS) have also been demonstrated to be effective for IFNβ-1b solubilization [93].…”

Section: Cell Lysis and Interferon Recoverymentioning

confidence: 99%

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Interferon-Based Biopharmaceuticals: Overview on the Production, Purification, and Formulation

et al. 2021

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The advent of biopharmaceuticals in modern medicine brought enormous benefits to the treatment of numerous human diseases and improved the well-being of many people worldwide. First introduced in the market in the early 1980s, the number of approved biopharmaceutical products has been steadily increasing, with therapeutic proteins, antibodies, and their derivatives accounting for most of the generated revenues. The success of pharmaceutical biotechnology is closely linked with remarkable developments in DNA recombinant technology, which has enabled the production of proteins with high specificity. Among promising biopharmaceuticals are interferons, first described by Isaacs and Lindenmann in 1957 and approved for clinical use in humans nearly thirty years later. Interferons are secreted autocrine and paracrine proteins, which by regulating several biochemical pathways have a spectrum of clinical effectiveness against viral infections, malignant diseases, and multiple sclerosis. Given their relevance and sustained market share, this review provides an overview on the evolution of interferon manufacture, comprising their production, purification, and formulation stages. Remarkable developments achieved in the last decades are herein discussed in three main sections: (i) an upstream stage, including genetically engineered genes, vectors, and hosts, and optimization of culture conditions (culture media, induction temperature, type and concentration of inducer, induction regimens, and scale); (ii) a downstream stage, focusing on single- and multiple-step chromatography, and emerging alternatives (e.g., aqueous two-phase systems); and (iii) formulation and delivery, providing an overview of improved bioactivities and extended half-lives and targeted delivery to the site of action. This review ends with an outlook and foreseeable prospects for underdeveloped aspects of biopharma research involving human interferons.

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Section: Clinical Importance Of Interferon-based Biopharmaceuticals and Market Overviewmentioning

confidence: 99%

Section: Therapeutic Cloned Interferonsmentioning

confidence: 99%

Section: Therapeutic Cloned Interferonsmentioning

confidence: 99%

Section: Therapeutic Cloned Interferonsmentioning

confidence: 99%

Section: Cell Lysis and Interferon Recoverymentioning

confidence: 99%

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Interferon-Based Biopharmaceuticals: Overview on the Production, Purification, and Formulation

et al. 2021

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Anthocyanin Partition in Aqueous Two‐Phase Systems Based on Isopropanol and Sodium/Ammonium Sulfate

et al. 2022

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Partitioning of anthocyanins contained in the crude extract of Syzygium cumini fruit was investigated in aqueous two‐phase systems (ATPS) made up of isopropanol and Na2SO4/(NH4)2SO4. Na2SO4 induced ATPS formation more effectively than (NH4)2SO4. The increase in temperature enhanced phase separation in the (NH4)2SO4 ATPS, while the opposite occurred in the Na2SO4 ATPS. The higher the overall mixture concentration or temperature, the higher the values of tie‐line length and slope. In all systems and conditions, anthocyanins preferentially partitioned to the top phase. The partition coefficient and theoretical recovery yield in the top phase varied in the ranges 1.14–1.77 and 53.31–63.87 % in the (NH4)2SO4 ATPS, and in the ranges 1.94–21.50 and 65.90–95.55 % in the Na2SO4 ATPS.

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Characterization of polymer‐polymer aqueous two‐phase system droplets for 3D culture future applications

Chairez‐Cantu

Rito‐Palomares

2023

J of Chemical Tech & Biotech

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BACKGROUND: Cell-based high-throughput platforms using aqueous two-phase systems (ATPS) have led to the construction of 3D cultures using ATPS in which stem cells are confined into polymer-polymer droplets, giving rise to viable spheroids for their further proliferation and differentiation. This work aims to characterize polymer-polymer ATPS droplet generation based on diameter size, morphology and uniformity, using basic laboratory equipment.RESULTS: Dextran (DEX), including DEX 500 000 and 70 000 g mol −1 , as well as UCON were used as the droplet phase (0.5, 1, 2 ∼L). Polyethylene glycol (PEG), including PEG 8 000, 10 000 and 35 000 g mol −1 , as well as DEX 75 000 g mol −1 and Ficoll were used as the bulk phase (50, 100 ∼L). Different ATPS compositions (molecular weights and concentrations) with phosphate-buffered saline as ATPS solvent and construction strategies (droplet added, immersed or covered) were evaluated in 96-well plates. ATPS droplets were generated in PEG-DEX ATPS. Diameter size and morphology were susceptible to ATPS compositions, construction strategies and DEX droplet phase volumes with no significant difference in PEG bulk phase volumes, whereas higher uniformity of ATPS droplets was correlated to lower PEG bulk volumes. Therefore, 1 ∼L DEX droplets immersed at the bottom of a well filled with 50 ∼L PEG resulted in uniform ATPS droplets with a mid-range diameter of 1.35 ± 0.23 mm. CONCLUSIONS: According to the results, this work demonstrates simplified and practical alternatives for the generation of reproducible PEG-DEX ATPS droplets that could be adapted for the encapsulation of different cell types for further 3D culture applications.

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Enhanced performance of polymer-polymer aqueous two-phase systems using ionic liquids as adjuvants towards the purification of recombinant proteins

Cited by 42 publications

References 61 publications

Interferon-Based Biopharmaceuticals: Overview on the Production, Purification, and Formulation

Interferon-Based Biopharmaceuticals: Overview on the Production, Purification, and Formulation

Anthocyanin Partition in Aqueous Two‐Phase Systems Based on Isopropanol and Sodium/Ammonium Sulfate

Characterization of polymer‐polymer aqueous two‐phase system droplets for 3D culture future applications

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