Performance of a pulsed-cap microcolumn for protein extraction

Abstract

-This work presents the results obtained using a microcolumn agitated by pulsed caps, using aqueous two-phase systems formed polyethylene glycol and salts (monobasic and dibasic) potassium phosphate for protein extraction. Proteins used were extracted cytochrome b5 and the enzyme ascorbic oxidoreductase. It was observed that operation of this equipment was stable and high efficiency values were achieved.

Extraction of Ascorbate Oxidase from Cucurbita maxima by Continuous Process in Perforated Rotating Disc Contactor Using Aqueous Two-Phase Systems

“…The results shown in Table 2 confirmed that PEG 20,000 (g/mol), PEG concentration of 20% (w/w), and citrate concentration of 15% (w/w) were the best conditions also for the purification factor (2.49). They are similar to those obtained for the extraction of AO in pulsed cap column (PEG 1,500/fosfato; P f =1.34) [8] and alpha toxin in PRDC using PEG/phosphate ATPS (P f =2.4) [7].…”

“…Liquid-liquid extraction equipments are widely used in biotechnology process for extraction of proteins [8,9], enzymes [8,[10][11][12], toxin [7], etc. The rotating disc contactor, consisting of a vertical shell equipped with a series of rotor discs and stators, can be used for this process [13].…”

Neural modeling for cytochrome b5 extraction

“…Experimental data were obtained by [1,2] with a batch extraction system and also with a continuous unit (column with pulsed caps). Both data sets have a maximum experimental deviation of AE10%.…”

“…Extracellular protease was produced by fermentation from MSD medium, at 30 8C during 96 h in a rotatory shaker. To avoid enzyme denaturation and loss of the main protein properties, a liquid-liquid extraction column with pulsed caps, which produces an efficient contact between the phases was used [1,2]. A batch extraction system was also studied.…”

A microdevice assisted approach for the preparation, characterization and selection of continuous aqueous two-phase systems: from micro to bench-scale

“…8 Since the driving forces for partitioning are of different nature and not completely understood, selecting an appropriate ATPS requires a complex experimental design, usually carried out in batch mode, which implies time and reagent waste. [9][10][11][12][13][14][15][16][17][18][19][20][21][22][23][24] Also, there is a lack of strategies to develop scaled and continuous processes that would allow the use of ATPS as a controlled and automated unit operation for biomolecule recovery at an industrial scale. Due to the great variety of phase components, compositions and biomolecule nature, a novel protocol or procedure that shortens data acquisition would be a great impulse to trigger scaling-up and industrial implementation of ATPS.…”