Aqueous two-phase systems: A viable platform in the manufacturing of biopharmaceuticals

Rosa, P.A.J.; Ferreira, Isabel; Azevedo, Ana M.; Aires-Barros, M. Raquel

doi:10.1016/j.chroma.2009.11.034

Cited by 225 publications

(179 citation statements)

References 49 publications

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“…Although chromatographic separations have been the workhorse of most purification processes, several limitations have been pointed out, such as (i) batch operation, (ii) low capacity, (iii) complex scale-up, (iv) time-consuming and high pressure packing processing, (v) slow intraparticle diffusion, (vi) low chemical and proteolytic stability and consequent contamination of the final product, and (vii) the high cost of the resins [5][6][7][8][9]. The replacement of some of these chromatographic steps by non-chromatographic alternatives, with high capacity and throughput, has been then suggested [7,[10][11][12].…”

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confidence: 99%

“…Aqueous two-phase extraction (ATPE) is a potential and promising non-chromatographic alternative, which can combine a high recovery, selectivity and biocompatibility with an easy scale-up and continuous operation mode [10,13,14]. Recently, the economic and environmental sustainability of an ATPE-based capture process for antibodies purification has been evaluated and compared to the currently established platform using protein A chromatography.…”

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confidence: 99%

“…Aqueous two-phase system (ATPS) has shown to be considerably advantageous in terms of process economy and operation, especially when processing high titer cell culture supernatants [13]. This liquid-liquid extraction (LLE) technology has been successfully reported in recent times as a primary stage unit operation in the downstream processing of several biological products including therapeutic proteins, such as antibodies [4,10,11,[15][16][17][18][19][20][21][22][23][24]. Its technical feasibility at large scale has been, however, demonstrated for the downstream processing of just few biological products [25][26][27].…”

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Continuous purification of antibodies from cell culture supernatant with aqueous two‐phase systems: From concept to process

Rosa

Azevedo

Sommerfeld

et al. 2013

Biotechnology Journal

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An aqueous two‐phase extraction (ATPE) process based on a PEG/phosphate system was developed for the capture of human immunoglobulin G and successfully applied to a Chinese hamster ovary and a PER.C6® cell supernatant. A continuous ATPE process incorporating three different steps (extraction, back‐extraction, and washing) was set up and validated in a pump mixer‐settler battery. Most of the higher molecular weight cell supernatant impurities were removed during the extraction step, while most of the lower molecular weight impurities were removed during the subsequent steps. A global recovery yield of 80% and a final protein purity of more than 99% were obtained for the IgG purification from a CHO cell supernatant, representing a 155‐fold reduction in the protein/IgG ratio. For the purification of IgG from a PER.C6® cell supernatant, a global recovery yield of 100%, and a host cell protein purity were attained, representing a 22‐fold reduction in the host cell protein/IgG ratio. These results, thus, open promising perspectives for the application of the developed ATPE process as a platform for the capture of antibodies. In fact, this new process has shown the ability to successfully recover and purify different antibodies from distinct cell culture supernatants. This technology can also overcome some of the limitations encountered using the typical chromatographic processes, besides inherent advantages of scalability, process integration, capability of continuous operation, and economic feasibility.

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Continuous purification of antibodies from cell culture supernatant with aqueous two‐phase systems: From concept to process

Rosa

Azevedo

Sommerfeld

et al. 2013

Biotechnology Journal

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“…Aqueous two-phase system (ATPS) extraction is a liquid-liquid extraction technique that has been used for the extraction and partial purification of a wide variety of biomolecules (Rosa et al 2010, Espitia-Saloma et al 2014, Reis et al 2014, Wu et al 2014. ATPS form when an aqueous solution exceeds a critical concentration of two water soluble, but mutually incompatible, components (Albertsson 1986).…”

Section: Introductionmentioning

confidence: 99%

Recovery of phenolic compounds from Eucalyptus wood wastes using ethanol-salt-based aqueous two-phase systems

Xavier¹,

Freire²,

Vidal³

2017

Maderas, Cienc. tecnol.

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In this work the partition behavior of phenols using ethanol-salt-based aqueous two phase systems (ATPS) was evaluated. The aim was the recovery of phenolic compounds with antioxidant capacity from eucalyptus wood industrial wastes. Experiments were planned to study the influence of several parameters on phenols partition, including type of inorganic salt (ammonium sulphate, a mixture of monopotassium phosphate and potassium diphosphate and potassium diphosphate), tie-lie lenght (TLL), volume ratio (V r ), settlement time and temperature. Phenols could be recovered preferently from the top or bottom phases depending on the salt used. It was demostrated that tie-lie lenght, volume ratio and temperature had influence on phenols partition. The highest total phenols yield 5,36 mg gallic acid equivalent (100 mg oven dried wood) -1 and FRAP antioxidant activity, 20 mmol AAE (100 g oven dried wood) -1 , was obtained using ATPS formed by 40.6% (w/w) ethanol and 12% (w/w) of ammonium sulphate at 65ºC. Analysis of the extract by RP-HPLC-ESI-TOF confirmed the presence of the phenolic compounds with potential antioxidant activity, namely, ellagic acid, myricetin 3-O-rhamnoside and quercetin 3-glucoside.

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“…In addition, the system can be easily scaled up. ATPS could be a good alternative for a first purification step since such systems allow removal of several contaminants by a simple and low-cost process (Albertsson 1986;Raghavarao et al 1995 andRajni 2001;Rosa et al 2010 and.…”

Section: Introductionmentioning

confidence: 99%

Aqueous two phase partitioning of fish proteins: partitioning studies and ATPS evaluation

Nagaraja

Iyyaswami

2014

J Food Sci Technol

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A suitable Aqueous Two Phase System (ATPS) has been identified for the partitioning of crude fish proteins from fish processing industrial effluent. A detailed study has been performed to analyze the influence of various parameters on protein partitioning such as molecular weight of PEG, effect of different salts (MgSO4 , K 2 HPO 4, Na 3 C 6 H 5 O 7, Na 2 SO 4, (NH 4 ) 2 PO 4, K 3 C 6 H 5 O 7 ) and their concentrations, pH, temperature, Tie Line Length (TLL), effluent loading and volume ratio. PEG 2000 -sodium sulphate ATPS was found to be a most favourable system among the selected ATPS for higher partition coefficient of proteins. The binodal curve and equilibrium characteristics of PEG 2000 -sodium sulphate were established and fitted to empirical equations. The equilibrium compositions (tie line) were correlated using Othmer-Tobias and Bancroft equations.

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Aqueous two-phase systems: A viable platform in the manufacturing of biopharmaceuticals

Cited by 225 publications

References 49 publications

Continuous purification of antibodies from cell culture supernatant with aqueous two‐phase systems: From concept to process

Continuous purification of antibodies from cell culture supernatant with aqueous two‐phase systems: From concept to process

Recovery of phenolic compounds from Eucalyptus wood wastes using ethanol-salt-based aqueous two-phase systems

Aqueous two phase partitioning of fish proteins: partitioning studies and ATPS evaluation

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