High-throughput screening-based selection and scale-up of aqueous two-phase systems for pDNA purification

2012 ◽  
Vol 35 (22) ◽  
pp. 3197-3207 ◽  
Author(s):  
Matthias Wiendahl ◽  
Stefan A. Oelmeier ◽  
Florian Dismer ◽  
Jürgen Hubbuch
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Scale Up ◽  
Two Phase ◽  
Aqueous Two Phase Systems ◽  
Phase Systems

scholarly journals High-throughput screening of aqueous two-phase systems for polyphenol extraction from A. nodosum: a green chemistry approach

2021 ◽  
Author(s):  
Alex Olivares-Molina ◽  
Brenda Parker
Keyword(s):  
Natural Products ◽  
Green Chemistry ◽  
High Throughput ◽  
High Throughput Screening ◽  
Scale Up ◽  
Model System ◽  
Two Phase ◽  
Aqueous Two Phase Systems ◽  
Scale Down ◽  
Phase Systems

Brown macroalgae are an attractive third-generation feedstock of natural products, in order to design green chemistry-compliant processes and reduce the use of organic solvents in bioactive product extraction, aqueous two-phase systems (ATPS) was applied. This research aimed to develop a high-throughput screening (HTS) to recover polyphenols from Ascophyllum nodosum using ATPS. In total, 384 different 2-phase systems were assessed using an automated liquid-handling system to evaluate polyphenol recovery using a model system of phloroglucinol to establish an optimal 2-phase system for polyphenol partitioning. Various ratios of PEG:potassium phosphate solutions were explored to evaluate partitioning of polyphenols via a scale-down approach. Scale-down selected system showed a recovery of phloroglucinol of 62.9±12.0%, this system was used for scale-up trials. Scale-up studies confirmed that the HTS method was able to recover polyphenols with a 54.8±14.2% in the phloroglucinol model system. When the optimised ATPS system was tested with a polyphenol extract, 93.62±8.24% recovery was observed. When ATPS was applied to a fucoidan and alginate biorefinery residue, 88.40±4.59% polyphenol was recovered. These findings confirm that ATPS is a valuable addition to the bioprocess toolkit for sustainable extraction of natural products from macroalgae in a multiproduct biorefinery approach.

High-Throughput 3D Neural Cell Culture Analysis Facilitated by Aqueous Two-Phase Systems

MRS Advances ◽  
2017 ◽  
Vol 2 (45) ◽  
pp. 2435-2441 ◽  
Author(s):  
Kristin Robin Ko ◽  
Rishima Agarwal ◽  
John Frampton
Keyword(s):  
Cell Culture ◽  
Cell Cultures ◽  
High Throughput ◽  
Hydroxypropyl Methylcellulose ◽  
3D Culture ◽  
Neural Cell ◽  
Epifluorescence Microscopy ◽  
Two Phase ◽  
Aqueous Two Phase Systems ◽  
Phase Systems

ABSTRACTThe three-dimensional (3D) culture of neural cells in extracellular matrix (ECM) gels holds promise for modeling neurodegenerative diseases and pre-clinical evaluation of novel therapeutics. However, most current strategies for fabricating 3D neural cell cultures are not well suited to automated production and analysis. Here, we present a facile, replicable, 3D cell culture system that is compatible with standard laboratory equipment and high-throughput workflows. This system uses aqueous two-phase systems (ATPSs) to confine small volumes (5 and 10 μl) of a commonly used ECM hydrogel (Matrigel) into thin, discrete layers, enabling highly-uniform production of 3D neural cell cultures in a 96-well plate format. These 3D neural cell cultures can be readily analyzed by epifluorescence microscopy and microplate reader. Our preliminary results show that many common polymers used in ATPSs interfere with Matrigel gelation and instead form fibrous precipitates. However, 0.5% hydroxypropyl methylcellulose (HPMC) and 2.5% dextran 10 kDa (D10) were observed to retain Matrigel integrity and had minimal impact on cell viability. This novel system offers a promising yet accessible platform for high-throughput fabrication of 3D neural tissues using readily available and cost-effective materials.

Aqueous two-phase systems as a tool for bioseparation – emphasis on organic acids

2020 ◽  
Vol 5 (9) ◽  
Author(s):  
Dragomir Yankov
Keyword(s):  
Organic Acids ◽  
Protein Separation ◽  
Large Scale ◽  
Process Integration ◽  
Scale Up ◽  
High Water ◽  
High Water Content ◽  
Two Phase ◽  
Aqueous Two Phase Systems ◽  
Phase Systems

AbstractAqueous two-phase systems (ATPS) are universally recognized as an excellent alternative to the conventional separation techniques in the biotechnology, because of their undoubted advantages such as mild and biocompatible conditions, high water content, low interfacial tension, ease of process integration and scale up, etc. The formation of ATPS is due to the incompatibility of two polymers in a common solution. Other types of ATPS are formed by polymer/salt, ionic and/or non-ionic surfactants, inorganic salt/short-chain alcohols, and based on room temperature ionic liquids. ATPS are successfully used (even in large scale) for cells, enzyme and protein separation, while their application for recovery of small molecules such as organic acids, antibiotics, alcohols is more complicated as they are usually hydrophilic and tend to distribute evenly between the phases. The purpose of this paper is to overview and summarize the efforts made for the application of different types of ATPS for the separation of organic acids.

High-throughput downstream process development for cell-based products using aqueous two-phase systems

2016 ◽  
Vol 1464 ◽  
pp. 1-11 ◽  
Author(s):  
Sarah Zimmermann ◽  
Sarah Gretzinger ◽  
Marie-Luise Schwab ◽  
Christian Scheeder ◽  
Philipp K. Zimmermann ◽  
...  
Keyword(s):  
High Throughput ◽  
Process Development ◽  
Downstream Process ◽  
Two Phase ◽  
Aqueous Two Phase Systems ◽  
Phase Systems

High-throughput downstream process development for cell-based products using aqueous two-phase systems (ATPS) - A case study

2016 ◽  
Vol 12 (2) ◽  
pp. 1600587 ◽  
Author(s):  
Sarah Zimmermann ◽  
Christian Scheeder ◽  
Philipp K Zimmermann ◽  
Are Bogsnes ◽  
Mattias Hansson ◽  
...  
Keyword(s):  
High Throughput ◽  
Process Development ◽  
Downstream Process ◽  
Two Phase ◽  
Aqueous Two Phase Systems ◽  
Phase Systems

scholarly journals Bioseparations in aqueous two-phase systems

2005 ◽  
pp. 145-154 ◽  
Author(s):  
Mirjana Antov
Keyword(s):  
Process Integration ◽  
Operation Mode ◽  
Scale Up ◽  
Operating Conditions ◽  
Two Phase ◽  
Environment Friendly ◽  
Aqueous Two Phase Systems ◽  
Low Interfacial Tension ◽  
Phase Systems ◽  
Rapid Mass

Bioseparations conducted in aqueous two-phase systems offer a great number of advantages over the conventional separation techniques. Among them the most relevant are rapid mass transfer due to low interfacial tension, rapid and selective separation, easiness of operation mode, reliability in scale-up biocompatibility and environment-friendly features, and possibility of process integration when applied in biomolecule production. Upon overcoming the major problem - mostly empirical establishment of operating conditions bioseparations in aqueous two-phase systems will become a necessary step in both existing and newly developed bioprocesses for the primary recovery of products.

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