High throughput screening techniques in downstream processing: Preparation, characterization and optimization of aqueous two-phase systems

2007 ◽  
Vol 62 (7) ◽  
pp. 2011-2021 ◽  
Author(s):  
Matthias Bensch ◽  
Björn Selbach ◽  
Jürgen Hubbuch
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Downstream Processing ◽  
Two Phase ◽  
Screening Techniques ◽  
Aqueous Two Phase Systems ◽  
Phase Systems

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.

scholarly journals Primary recovery of hyaluronic acid produced in Streptococcus equi subsp. zooepidemicus using PEG–citrate aqueous two-phase systems

AMB Express ◽  
2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Miguel Flores-Gatica ◽  
Héctor Castañeda-Aponte ◽  
Mónica Rebeca Gil-Garzon ◽  
Liliana Monserrath Mora-Galvez ◽  
Martin Paul Banda-Magaña ◽  
...  
Keyword(s):  
Hyaluronic Acid ◽  
Negative Impact ◽  
Fermentation Broth ◽  
Downstream Processing ◽  
Bottom Phase ◽  
Two Phase ◽  
Streptococcus Equi ◽  
Aqueous Two Phase Systems ◽  
Phase Systems ◽  
First Time

AbstractGiven its biocompatibility, rheological, and physiological properties, hyaluronic acid (HA) has become a biomaterial of increasing interest with multiple applications in medicine and cosmetics. In recent decades, microbial fermentations have become an important source for the industrial production of HA. However, due to its final applications, microbial HA must undergo critical and long purification processes to ensure clinical and cosmetic grade purity. Aqueous two-phase systems (ATPS) have proven to be an efficient technique for the primary recovery of high-value biomolecules. Nevertheless, their implementation in HA downstream processing has been practically unexplored. In this work, polyethylene glycol (PEG)–citrate ATPS were used for the first time for the primary recovery of HA produced with an engineered strain of Streptococcus equi subsp. zooepidemicus. The effects of PEG molecular weight (MW), tie-line length (TLL), volume ratio (VR), and sample load on HA recovery and purity were studied with a clarified fermentation broth as feed material. HA was recovered in the salt-rich bottom phase, and its recovery increased when a PEG MW of 8000 g mol−1 was used. Lower VR values (0.38) favoured HA recovery, whereas purity was enhanced by a high VR (3.50). Meanwhile, sample load had a negative impact on both recovery and purity. The ATPS with the best performance was PEG 8000 g mol−1, TLL 43% (w/w), and VR 3.50, showing 79.4% HA recovery and 74.5% purity. This study demonstrated for the first time the potential of PEG–citrate ATPS as an effective primary recovery strategy for the downstream process of microbial HA.

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.

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

Downstream processing of virus, virus-like particles and nanoparticulate inclusion bodies to be used as gene delivery vehicles for human gene therapy applications

2021 ◽  
Author(s):  
Moataz Dowaidar
Keyword(s):  
Environmental Sustainability ◽  
Inclusion Bodies ◽  
Culture Media ◽  
Downstream Processing ◽  
Economic Feasibility ◽  
Two Phase ◽  
Affinity Ligands ◽  
Virus Like Particles ◽  
Aqueous Two Phase Systems ◽  
Phase Systems

Design and analyze future affinity ligands such as antibodies, aptamers, metal affinity, peptides, triazine dyes, and red blood cell carriers to increase monolithic chromatography selectivity and two-phase affinity-aqueous systems. Monolithic chromatography systems may be constructed and optimized by examining viruses, virus-like particles, and nanoparticulate inclusion bodies from difficult culture media. Different chromatographic characteristics such as monolithic supports, active monolith groups, binding and elution buffers, elution gradients, column equilibrium volumes, and regeneration should be investigated with the aim of optimizing monolithic chromatography separation and recovery. To develop and optimize aqueous two-phase systems, several different parameters need to be investigated and compared, such as polymer type and concentration, salt solution, surfactants and ions, polymer molecular weight, ion nature and ionic strength, present affinity ligands, volumetric ratio, temperature, pH, and sample load, to name a few.Several relevant biological models, such as Newcastle disease (NDV), herpes (HPr), or viral vectors bluetongue (BTV), can be employed to establish different patterns and predict the generalized mechanistic process of enhanced purification systems. Because optimal systems are intended to be deployed at the industrial level, a comparison of typical downstream processing and alternatives in terms of recovery, technological advantages, economic feasibility, and environmental sustainability is needed. Cell separation, viral concentration and viral purification may all be reduced to one step utilizing integrated chromatography platforms on monolith supports and aqueous two-phase systems. In addition, the efficiency of separation and purification yields achieved after these integrated systems are predicted to be substantially higher than those produced in conventional procedures. As a result, the benefits of these alternative and cutting-edge technologies are predicted to contribute to more efficient, long-term, and cost-effective downstream processing of virus-like particles.

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