Compared to other integrating viral vectors, foamy virus (FV) vectors have distinct advantages as a gene transfer tool, including their nonpathogenicity, the ability to carry larger transgene cassettes, and increased stability of virus particles due to DNA genome formation within the virions. Proof of principle of its therapeutic utility was provided with the correction of canine leukocyte adhesion deficiency using autologous CD34 cells transduced with FV vector carrying the canine CD18 gene, demonstrating its long-term safety and efficacy. However, infectious titers of FV-human(h)CD18 were low and not suitable for manufacturing of clinical-grade product. Herein, we developed a scalable production and purification process that resulted in 60-fold higher FV-hCD18 titers from ~1.7 × 10 to 1.0 × 10 infectious units (IU)/ml. Process development improvements included use of polyethylenimine-based transfection, use of a codon-optimized , heparin affinity chromatography, tangential flow filtration, and ultracentrifugation, which reproducibly resulted in 5,000-fold concentrated and purified virus, an overall yield of 19 ± 3%, and final titers of 1-2 × 10 IU/ml. Highly concentrated vector allowed reduction of final dimethyl sulfoxide (DMSO) concentration, thereby avoiding DMSO-induced toxicity to CD34 cells while maintaining high transduction efficiencies. This process development results in clinically relevant, high titer FV which can be scaled up for clinical grade production.
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http://dx.doi.org/10.1038/mtm.2016.4 | DOI Listing |
PLoS One
December 2024
Department of Water Resources, Climatology and Environmental Management, University of Warmia and Mazury in Olsztyn, Olsztyn, Poland.
Introduction: Lung cancer, one of the leading causes of death due to neoplasms, requires prompt diagnosis and immediate treatment. The COVID-19 pandemic affected healthcare systems worldwide, having adverse effects on all aspects, particularly on the fate of patients with suspected neoplastic diseases. Limited access to healthcare, disruptions in regular operations (reassigning roles to some wards), postponed hospital admissions, prolonged diagnostic processes, and other factors have collectively led to the phenomenon known as COVID-19 debt.
View Article and Find Full Text PDFMethods Mol Biol
December 2024
Bioprotection Aotearoa, School of Food Technology and Natural Sciences, Massey University, Palmerston North, New Zealand.
This chapter describes the protocol for heterologous expression of Phytophthora proteins in the yeast Pichia pastoris. Two methods to prepare the constructs for expression are described, using two different strains of P. pastoris, as well as methods for protein expression and purification by immobilized metal ion affinity (IMAC).
View Article and Find Full Text PDFToxins (Basel)
December 2024
Environmental Technology and Water Resources Postgraduate Program, Department of Civil and Environmental Engineering, University of Brasília, Brasília 70910-900, Brazil.
The frequency and intensity of harmful cyanobacterial blooms have increased in the last decades, posing a risk to public health since conventional water treatments do not effectively remove extracellular cyanotoxins. Consequently, advanced technologies such as the Fenton process are required to ensure water safety. The cyanotoxin cylindrospermopsin (CYN) demands special attention, as it is abundant in the extracellular fraction and has a high toxicological potential.
View Article and Find Full Text PDFToxins (Basel)
December 2024
CEA, INRAE, Département Médicaments et Technologies pour la Santé (DMTS), SPI, Université Paris-Saclay, 91191 Gif-sur-Yvette, France.
Food poisoning outbreaks frequently involve staphylococcal enterotoxins (SEs). SEs include 33 distinct types and multiple sequence variants per SE type. Various mass spectrometry methods have been reported for the detection of SEs using a conventional bottom-up approach.
View Article and Find Full Text PDFNanomaterials (Basel)
December 2024
Laboratory of Advanced Materials and Technology, Tomsk State University, Tomsk 634050, Russia.
Photocatalysis offers a powerful approach for water purification from toxic organics, hydrogen production, biosolids processing, and the conversion of CO into useful products. Further advancements in photocatalytic technologies depend on the development of novel, highly efficient catalysts and optimized synthesis methods. This study aimed to develop a laser synthesis technique for bismuth oxyhalide nanoparticles (NPs) as efficient and multifunctional photocatalysts.
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