Removal of proteases from Clostridium perfringens fermented broth by aqueous two-phase systems (PEG/citrate)

Porto, Tatiana Souza; Pessôa-Filho, Pedro Alcântara; Neto, B.B.; Filho, J. L. Lima; Converti, Attílio; Porto, Ana Lúcia Figueiredo; Pessoa, Adalberto

doi:10.1007/s10295-007-0230-8

Cited by 31 publications

(17 citation statements)

References 26 publications

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“…In order to establish the conditions under which the phase separation occurs, binodal curves, formed by PEG and sodium citrate, were investigated at different values of pH and PEG molar mass. After statistical experimental planning, it was found that simultaneous increase in PEG molar mass and decrease in citrate concentration remarkably improved the removal factor, whereas the protease removal yield showed an opposite trend, 50 confirming previous results reported by Porto and co-workers. 49 Finally, bromelain (enzymatic complex obtained from pineapple, Ananas comosus, fruits and stem) was investigated by thermoseparation using by poly(ethylene oxide) (PEO)-poly(propylene oxide) (PPO)-poly (ethylene oxide) (PEO)block copolymers aqueous solutions.…”

Section: Aqueous Two-phase Polymeric Systems -Atppssupporting

confidence: 87%

Liquid–liquid extraction of biomolecules: an overview and update of the main techniques

Mazzola

Lopes

Hasmann

et al. 2007

J of Chemical Tech & Biotech

226

119

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Two-phase systems can be exploited in separation science for the extraction/purification of desired biomolecules. This article reviews recent experimental work on the use of aqueous two-phase polymer systems, two-phase aqueous micellar systems and reversed micellar systems for the extraction/purification of biomolecules. The experimental partitioning behavior of different biomolecules is reviewed and new results with nisin and lipopolysaccharides are presented. An extensive description of each system is presented, based on the biomolecules behavior and the tools employed to improve their extraction and purification.

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Section: Aqueous Two-phase Polymeric Systems -Atppssupporting

confidence: 87%

Liquid–liquid extraction of biomolecules: an overview and update of the main techniques

Mazzola

Lopes

Hasmann

et al. 2007

J of Chemical Tech & Biotech

226

119

View full text Add to dashboard Cite

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“…The observation that adenovirus particles partitioned and concentrated preferably to the top phase can be explained by the excluded volume effect , common in high mass molecules partition in these systems . Adenoviral particles have large diameters (approximately 100 nm ), and a greater exclusion effect seems to occurs in TX‐114 abundant systems in our results as schematized in Fig.…”

Section: Resultsmentioning

confidence: 53%

Application of aqueous two‐phase micellar system to improve extraction of adenoviral particles from cell lysate

Molino

Lopes

Marques

et al. 2017

Biotech and App Biochem

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Viral vectors are important in medical approaches, such as disease prevention and gene therapy, and their production depends on efficient prepurification steps. In the present study, an aqueous two-phase micellar system (ATPMS) was evaluated to extract human adenovirus type 5 particles from a cell lysate. Adenovirus was cultured in human embryonic kidney 293 (HEK-293) cells to a concentration of 1.4 × 10 particles/mL. Cells were lysed, and the system formed by direct addition of Triton X-114 in a 2 full factorial design with center points. The systems were formed with Triton X-114 at a final concentration of 1.0, 6.0, and 11.0% (w/w), cell lysate pH of 6.0, 6.5, and 7.0, and incubation temperatures at 33, 35, and 37 °C. Adenovirus particles recovered from partition phases were measured by qPCR. The best system condition was with 11.0% (w/w) of Triton X-114, a cell lysate pH of 7.0, and an incubation temperature at 33 °C, yielding 3.51 × 10 adenovirus particles/mL, which increased the initial adenovirus particles concentration by 2.3-fold, purifying it by 2.2-fold from the cell lysate, and removing cell debris. In conclusion, these results demonstrated that the use of an aqueous two-phase micellar system in the early steps of downstream processing could improve viral particle extraction from cultured cells while integrating clarification, concentration, and prepurification steps.

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“…From E. coli fermented broth or cell lysate, the extraction of proteins, virus, plasmid DNA (pDNA) and enzymes has been reported . ABS‐based protocols can also be applied to recover toxin A and enzymes, from Clostridium perfringens , virus from Salmonella typhimurium, pigments from Spirulina platensis, enzymes from Penicillium restrictum, Penicillium janthinellum , and Bacillus sp, antibiotics from Streptomyces aureofaciens and pharmaceuticals from Streptomyces clavuligerus , . Yeast is also popular for biomolecules synthesis, and there are ABS studies showing the recovery of enzymes from S. cerevisiae and proteins from P. pastoris and bakers' yeast …”

Section: Introductionmentioning

confidence: 99%

Liquid–liquid extraction of biopharmaceuticals from fermented broth: trends and future prospects

Santos

Santos-Ebinuma

Pessoa

et al. 2017

J of Chemical Tech & Biotech

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Biopharmaceuticals are one of the most important groups of biotechnological products, representing one-quarter of all pharmaceutical sales and providing suitable and efficient medical care for many previously untreatable diseases. However, their production and purification usually require complex processes, resulting in extremely expensive final products. Thus, the development of newer, simpler and cheaper methods for biosynthesis and purification of these biocompounds, as well as proper integration between these stages, are crucial to reduce their commercial costs and to increase the scale of biopharmaceuticals' medical use. One solution for this concern relies on the proper integration between upstream and downstream processing applying liquid-liquid extraction systems as an intermediate stage. Recently, several works reported that a proper choice of liquid-liquid systems such as aqueous biphasic systems (ABS) can be a suitable platform to simplify, reduce the number of the downstream stages, improve the recovery and purification yields and, consequently, decrease biopharmaceuticals' manufacturing costs. This review will explain the general concept of biopharmaceuticals, their main production and purification methods, particularly, exploring the use of ABS as effective and integrative platforms for their recovery and purification, and providing further insights into the future trends and prospects in the field.

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Removal of proteases from Clostridium perfringens fermented broth by aqueous two-phase systems (PEG/citrate)

Cited by 31 publications

References 26 publications

Liquid–liquid extraction of biomolecules: an overview and update of the main techniques

Liquid–liquid extraction of biomolecules: an overview and update of the main techniques

Application of aqueous two‐phase micellar system to improve extraction of adenoviral particles from cell lysate

Liquid–liquid extraction of biopharmaceuticals from fermented broth: trends and future prospects

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