Optimization and miniaturization of aqueous two phase systems for the purification of recombinant human immunodeficiency virus-like particles from a CHO cell supernatant

Jacinto, Maria João; Soares, Ruben R. G.; Azevedo, Ana M.; Chu, V.; Tover, Andres; Conde, J.P.; Aires-Barros, M. Raquel

doi:10.1016/j.seppur.2015.09.006

Cited by 46 publications

(35 citation statements)

References 41 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Jacinto et al optimized an ATPS for the purification of human immunodeficiency virus‐like particles at mL‐scale and successfully achieved similar results in a microfluidic device. ATPSs composed of PEG/salts and PEG/dextran were investigated and the highest K obtained was for the PEG/ammonium sulfate system that had a partition value of 3.9, comparable to the K obtained in macroscale for the same system ( K = 4.4), thus validating the microfluidic approach as a potential screening tool .…”

Section: Current and Emerging Microfluidic Atps Platformssupporting

confidence: 55%

Miniaturization of aqueous two‐phase extraction for biological applications: From micro‐tubes to microchannels

Soares

Silva

Fernandes

et al. 2016

Biotechnology Journal

View full text Add to dashboard Cite

Aqueous two-phase extraction (ATPE) is a biocompatible liquid-liquid (L-L) separation technique that has been under research for several decades towards the purification of biomolecules, ranging from small metabolites to large animal cells. More recently, with the emergence of rapid-prototyping techniques for fabrication of microfluidic structures with intricate designs, ATPE gained an expanded range of applications utilizing physical phenomena occurring exclusively at the microscale. Today, research is being carried simultaneously in two different volume ranges, mL-scale (microtubes) and nL-scale (microchannels). The objective of this review is to give insight into the state of the art at both microtube and microchannel-scale and to analyze whether miniaturization is currently a competing or divergent technology in a field of applications including bioseparation, bioanalytics, enhanced fermentation processes, catalysis, high-throughput screening and physical/chemical compartmentalization. From our perspective, both approaches are worthy of investigation and, depending on the application, it is likely that either (i) one of the approaches will eventually become obsolete in particular research areas such as purification at the preparative scale or high-throughput screening applications; or (ii) both approaches will function as complementing techniques within the bioanalytics field.

show abstract

Section: Current and Emerging Microfluidic Atps Platformssupporting

confidence: 55%

Miniaturization of aqueous two‐phase extraction for biological applications: From micro‐tubes to microchannels

Soares

Silva

Fernandes

et al. 2016

Biotechnology Journal

View full text Add to dashboard Cite

show abstract

“…The importance of viral particles in biotechnology has increased in recent years as they have been used in applications including molecular library screening, drug delivery, cancer treatment, diagnostics, vaccination, control of foodborne pathogens, gene therapy and bioimaging . Currently, M13 has been extensively used in phage display technology, bionanomaterial construction as well as biosensors development …”

Section: Introductionmentioning

confidence: 99%

“…ATPS have been exploited for several years for the recovery of bioproducts such as proteins, cells, organelles, genetic material, low‐molecular weight metabolites and viral particles . Concerning virus recovery, ATPS have been proposed for the recovery of rotavirus‐like particles, bacteriophage T4, adenovirus, porcine parvovirus, B19 parvovirus‐like particles and recombinant human immunodeficiency virus‐like particles . Application of ATPS to viruses is not common, particularly when compared with reports addressing the recovery of proteins and other types of macromolecules.…”

Section: Introductionmentioning

confidence: 99%

Recovery and primary purification of bacteriophage M13 using aqueous two‐phase systems

González‐Mora

Ruiz‐Ruiz

Benavides

et al. 2017

J of Chemical Tech & Biotech

View full text Add to dashboard Cite

BACKGROUND Bacteriophage M13 is an Escherichia coli‐specific non‐lytic filamentous virus commonly used in applications ranging from antibody screening and nanomaterial construction to drug delivery, among others. In this tenor, alternative methods for the fractionation, recovery and partial purification of phage particles are desired. In this work, the use of aqueous two‐phase systems (ATPS) was evaluated as an alternative method for the recovery of phage particles. RESULTS The partition behavior of M13 in PEG–salt and ionic liquid (IL)–salt ATPS was characterized using a pre‐purified feedstock. In PEG‐salt ATPS, M13 was preferentially partitioned to the interface. In IL ATPS, however, M13 showed a high‐top phase preference with recovery yields above 65%. Selected systems were tested for the extraction of M13 from a crude fermentation broth. From crude broth, a PEG 400‐potassium phosphate system with volume ratio (VR) of 1 and 25% w/w tie line length (TLL) gave the best M13 top phase recovery (83%) and purification fold (18.2) in terms of total protein concentration. CONCLUSIONS The results presented here demonstrate the practical application of ATPS as an efficient process for the primary recovery and partial purification of M13 and represent the first study of the extraction of viral particles directly from a crude broth as well as the use of IL‐Salt ATPS. © 2017 Society of Chemical Industry

show abstract

“…Benavides et al used PEG–potassium phosphate system for the recovery and primary purification of double layered Rotavirus‐like particles (dlRLP) produced using baculovirus expression system . Jacinto et al reported on ABS purification of recombinant HIV‐VLPs from a cell supernatant, and Lueachau et al studied the recovery of B19 virus‐like particles using PEG 1000‐magnesium sulfate ABS . However, until now the use of ILs in implementation of ABS to improve VLPs separation and purification has not been reported.…”

Section: Introductionmentioning

confidence: 99%

High‐throughput screening of aqueous biphasic systems with ionic liquids as additives for extraction and purification of enveloped virus‐like particles

Marchel

Soares

Vormittag

et al. 2019

Engineering Reports

View full text Add to dashboard Cite

Due to their unique properties, virus‐like particles (VLPs) have been portrayed as a promising high‐value biopharmaceutical in VLP‐based vaccination and cancer therapy. Nevertheless, due to limited physical and economical capabilities of the current downstream processing of VLPs, their production is still difficult and seen as a major problem that needs to be tackled. In this work, high‐throughput screening on a liquid handling station (LHS) has been implemented for efficient selection of adequate polymer‐salt aqueous biphasic systems (ABS) for extraction and purification of enveloped Hepatitis C virus pseudoparticles (HCVpp). The effect of polyethylene glycol (PEG) molecular weight and salt type (citrate, sulfate, and phosphate) was first evaluated. Furthermore, to optimize extraction parameters, the effect of pH and tie‐line lengths (TLL) was also addressed. For the most promising ABS and extraction conditions, the addition of ten ionic liquids (ILs) as adjuvants was investigated. Insights on the chemical features of ILs that impact HCV‐VLPs partitioning are highlighted. Finally, the potential of studied ABS containing ILs as adjuvants in the extraction and purification of VLPs from cell culture supernatants was addressed. The 100% extraction efficiency of VLPs in the PEG‐rich phase was achieved, and VLP purity was increased compared to the system without IL. In conclusion, these promising results show that ILs can be very effective in modulating the phase properties of polymer‐salt ABS, achieving high HCV‐VLP purification.

show abstract

Optimization and miniaturization of aqueous two phase systems for the purification of recombinant human immunodeficiency virus-like particles from a CHO cell supernatant

Cited by 46 publications

References 41 publications

Miniaturization of aqueous two‐phase extraction for biological applications: From micro‐tubes to microchannels

Miniaturization of aqueous two‐phase extraction for biological applications: From micro‐tubes to microchannels

Recovery and primary purification of bacteriophage M13 using aqueous two‐phase systems

High‐throughput screening of aqueous biphasic systems with ionic liquids as additives for extraction and purification of enveloped virus‐like particles

Contact Info

Product

Resources

About