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https://www.readbyqxmd.com/read/29152858/in-vitro-evaluation-of-anti-aggregation-and-degradation-behavior-of-pegylated-polymeric-nanogels-under-in-vivo-like-conditions
#1
Yinan Chen, George R Dakwar, Kevin Braeckmans, Twan Lammers, Wim E Hennink, Josbert M Metselaar
The in vivo stability and biodegradability of nanocarriers crucially determine therapeutic efficacy as well as safety when used for drug delivery. This study aims to evaluate optimized in vitro techniques predictive for in vivo nanocarrier behavior. Polymeric biodegradable nanogels based on hydroxyethyl methacrylamide-oligoglycolates-derivatized poly(hydroxyethyl methacrylamide-co-N-(2-azidoethyl)methacrylamide) and with various degrees of PEGylation and crosslinking densities are prepared. Three techniques are chosen and refined for specific in vitro evaluation of the nanocarrier performance: (1) fluorescence single particle tracking (fSPT) to study the stability of nanogels in human plasma, (2) tangential flow filtration (TFF) to study the degradation and filtration of nanogel degradation products, and (3) fluorescence fluctuation spectroscopy (FFS) to evaluate and compare the degradation behavior of nanogels in buffer and plasma...
November 20, 2017: Macromolecular Bioscience
https://www.readbyqxmd.com/read/29076623/inflammation-stimulated-mesenchymal-stromal-cell-derived-extracellular-vesicles-attenuate-inflammation
#2
Matthew T Harting, Amit K Srivastava, Siqin Zhaorigetu, Henry Bair, Karthik S Prabhakara, Naama E Toledano Furman, Jody V Vykoukal, Katherine A Ruppert, Charles S Cox, Scott D Olson
Extracellular vesicles (EVs) secreted by mesenchymal stromal cells (MSCs) have been proposed to be a key mechanistic link in the therapeutic efficacy of cells in response to cellular injuries through paracrine effects. We hypothesize that inflammatory stimulation of MSCs results in the release of EVs that have greater anti-inflammatory effects. The present study evaluates the immunomodulatory abilities of EVs derived from inflammation-stimulated and naive MSCs (MSCEv(+) and MSCEv, respectively) isolated using a current Good Manufacturing Practice (cGMP)-compliant tangential flow filtration (TFF) system...
October 27, 2017: Stem Cells
https://www.readbyqxmd.com/read/29043350/microfluidic-diafiltration-on-chip-using-an-integrated-magnetic-peristaltic-micropump
#3
Jessica F Liu, Sagar Yadavali, Andrew Tsourkas, David Issadore
Diafiltration is a membrane filtration technique that rapidly removes permeable molecules from a solution by controlling the tangential and orthogonal flows over a membrane and by replenishing the permeate with an equivalent amount of replacement buffer. However, its application to the purification of many key biomaterials and nanomaterials has been limited by the large dead volume (>10 mL) that is required to automate the process. To address this challenge, we have developed a diafiltration-on-a-chip device that can process low-volume samples (50 μL)...
November 7, 2017: Lab on a Chip
https://www.readbyqxmd.com/read/28992119/a-reproducible-and-scalable-procedure-for-preparing-bacterial-extracts-for-cell-free-protein-synthesis
#4
Kazushige Katsura, Takayoshi Matsuda, Yuri Tomabechi, Mayumi Yonemochi, Kazuharu Hanada, Noboru Ohsawa, Kensaku Sakamoto, Chie Takemoto, Mikako Shirouzu
Cell-free protein synthesis is a useful method for preparing proteins for functional or structural analyses. However, batch-to-batch variability with regard to protein synthesis activity remains a problem for large-scale production of cell extract in the laboratory. To address this issue, we have developed a novel procedure for large-scale preparation of bacterial cell extract with high protein synthesis activity. The developed procedure comprises cell cultivation using a fermentor, harvesting and washing of cells by tangential flow filtration, cell disruption with high-pressure homogenizer and continuous diafiltration...
November 1, 2017: Journal of Biochemistry
https://www.readbyqxmd.com/read/28840656/effect-of-zinc-chloride-and-peg-concentrations-on-the-critical-flux-during-tangential-flow-microfiltration-of-bsa-precipitates
#5
Zhao Li, Andrew L Zydney
There is renewed interest in the possibility of using precipitation for initial capture of high value therapeutic proteins as part of an integrated continuous downstream process. These precipitates can be continuously washed using tangential flow filtration, with long term operation achieved by operating the membrane modules below the critical filtrate flux for fouling. Our hypothesis was that the critical flux for the precipitated protein would be a function of the properties of the precipitate as determined by the precipitation conditions...
August 25, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28828646/sequential-filtration-a-gentle-method-for-the-isolation-of-functional-extracellular-vesicles
#6
Mitja L Heinemann, Jody Vykoukal
A prevalent challenge in isolating extracellular vesicles (EVs) from biological fluids is the reliable depletion of abundant contaminants-including free proteins and biomolecules, as well as nontarget vesicle subpopulations and other nanoparticulates-from the sample matrix while maximizing recovery. Sequential Filtration is a recently published approach for the size-based isolation of exosomes that is ideally suited for large-volume biofluid samples such as ascites , urine , lavage fluid, or cell-conditioned media...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28776064/manufacturing-of-proteins-and-antibodies-chapter-downstream-processing-technologies-harvest-operations
#7
Richard Turner, Adrian Joseph, Nigel Titchener-Hooker, Jean Bender
Cell harvesting is the separation or retention of cells and cellular debris from the supernatant containing the target molecule Selection of harvest method strongly depends on the type of cells, mode of bioreactor operation, process scale, and characteristics of the product and cell culture fluid. Most traditional harvesting methods use some form of filtration, centrifugation, or a combination of both for cell separation and/or retention. Filtration methods include normal flow depth filtration and tangential flow microfiltration...
August 4, 2017: Advances in Biochemical Engineering/biotechnology
https://www.readbyqxmd.com/read/28751635/microfluidic-cell-retention-device-for-perfusion-of-mammalian-suspension-culture
#8
Taehong Kwon, Holly Prentice, Jonas De Oliveira, Nyasha Madziva, Majid Ebrahimi Warkiani, Jean-François P Hamel, Jongyoon Han
Continuous production of biologics, a growing trend in the biopharmaceutical industry, requires a reliable and efficient cell retention device that also maintains cell viability. Current filtration methods, such as tangential flow filtration using hollow-fiber membranes, suffer from membrane fouling, leading to significant reliability and productivity issues such as low cell viability, product retention, and an increased contamination risk associated with filter replacement. We introduce a novel cell retention device based on inertial sorting for perfusion culture of suspended mammalian cells...
July 27, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28648546/process-intensification-for-high-yield-production-of-influenza-h1n1-gag-virus-like-particles-using-an-inducible-hek-293-stable-cell-line
#9
Alina Venereo-Sanchez, Melanie Simoneau, Stéphane Lanthier, Parminder Chahal, Lucie Bourget, Sven Ansorge, Rénald Gilbert, Olivier Henry, Amine Kamen
Influenza virus dominant antigens presentation using virus like particle (VLP) approach is attractive for the development of new generation of influenza vaccines. Mammalian cell platform offers many advantages for VLP production. However, limited attention has been paid to the processing of mammalian cell produced VLPs. Better understanding of the production system could contribute to increasing the yields and making large-scale VLP vaccine manufacturing feasible. In a previous study, we have generated a human embryonic kidney HEK-293 inducible cell line expressing Hemagglutinin (HA) and Neuraminidase (NA), which was used to produce VLPs upon transient transfection with a plasmid containing HIV-1 Gag...
June 22, 2017: Vaccine
https://www.readbyqxmd.com/read/28645718/optimizing-adjuvants-for-intradermal-delivery-of-menc-glycoconjugate-vaccine
#10
Agnese Donadei, Cristiana Balocchi, Maria R Romano, Luigi Panza, Roberto Adamo, Francesco Berti, Derek T O'Hagan, Simona Gallorini, Barbara C Baudner
Intradermal vaccine delivery is a promising alternative to the conventional intramuscular route. The skin layer is immunologically supported by a densely network of antigen presenting cells, while the skeletal muscle is loaded with a relatively sparse population of APCs. Nevertheless, the vaccine to be suitable for intradermal delivery needs a new formulation to facilitate either smaller injection volumes or the introduction into new delivery devises as micro-needles. This study presents a proof of concept for intradermal delivery of the MenC-CRM197 glycoconjugate vaccine using a mouse model...
July 13, 2017: Vaccine
https://www.readbyqxmd.com/read/28480535/integrated-continuous-bioprocessing-economic-operational-and-environmental-feasibility-for-clinical-and-commercial-antibody-manufacture
#11
James Pollock, Jon Coffman, Sa V Ho, Suzanne S Farid
This paper presents a systems approach to evaluating the potential of integrated continuous bioprocessing for monoclonal antibody (mAb) manufacture across a product's lifecycle from preclinical to commercial manufacture. The economic, operational, and environmental feasibility of alternative continuous manufacturing strategies were evaluated holistically using a prototype UCL decisional tool that integrated process economics, discrete-event simulation, environmental impact analysis, operational risk analysis, and multiattribute decision-making...
July 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28464237/ultrafiltration-behavior-of-monoclonal-antibodies-and-fc-fusion-proteins-effects-of-physical-properties
#12
Youngbin Baek, Nripen Singh, Abhiram Arunkumar, Michael Borys, Zheng J Li, Andrew L Zydney
Ultrafiltration (UF) is used for the final concentration and formulation of essentially all antibody-based therapeutics including both monoclonal antibodies (mAbs) and Fc-fusion proteins. The objective of this study was to quantitatively compare the filtrate flux behavior for two highly purified mAbs and an Fc-fusion protein under identical flow and buffer conditions. Filtrate flux data were obtained using a Pellicon 3 tangential flow filtration cassette over a wide range of transmembrane pressures and bulk protein concentrations...
September 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28440049/analytical-based-methodologies-for-examining-the-in-vitro-absorption-distribution-metabolism-and-elimination-adme-of-silver-nanoparticles
#13
Sesha L A Paluri, John D Ryan, Nhi H Lam, Dhriti Nepal, Ioana E Sizemore
The clinical applications of silver nanoparticles (AgNPs) remain limited due to the lack of well-established methodologies for studying their nanokinetics. Hereby, the primary goal is to adapt a suite of analytical-based methodologies for examining the in vitro absorption, distribution, metabolism, and elimination of AgNPs. Vero 76 and HEK 293 cells are exposed to ≈10-nm spherical AgNPs(+) and AgNPs(-) at relevant concentrations (0-300 µg mL(-1) ) and times (4-48 h). Absorption: Inductively coupled plasma optical emission spectroscopy (ICP-OES) demonstrates that the two AgNP formulations are not bioequivalent...
April 25, 2017: Small
https://www.readbyqxmd.com/read/28403748/an-effective-method-of-atelocollagen-type-1-3-isolation-from-human-placenta-and-its-in-vitro-characterization-in-two-dimesional-and-three-dimensional-cell-culture-applications
#14
Johannes Hackethal, Severin Mühleder, Alexandra Hofer, Karl Heinrich Schneider, Johanna Prüller, Simone Hennerbichler, Heinz Redl, Andreas Teuschl
Pepsin-solubilized atelocollagen can be used to form highly complex three-dimensional matrices for a broad spectrum of tissue engineering applications. Moreover, it has a long history as a favorable biomaterial in pharmaceutical and medical industries. So far, the main sources for these approaches are collagens from xenogenic sources. Yet, these nonhuman collagens carry a risk of provoking immune reactions in patients. Here we describe an effective method of isolating atelocollagen type 1/3 (COL1/3) from human placenta...
May 2017: Tissue Engineering. Part C, Methods
https://www.readbyqxmd.com/read/28395932/microalgae-fractionation-using-steam-explosion-dynamic-and-tangential-cross-flow-membrane-filtration
#15
E Lorente, M Hapońska, E Clavero, C Torras, J Salvadó
In this study, the microalga Nannochloropsis gaditana was subjected to acid catalysed steam explosion treatment and the resulting exploded material was subsequently fractionated to separate the different fractions (lipids, sugars and solids). Conventional and vibrational membrane setups were used with several polymeric commercial membranes. Two different routes were followed: 1) filtration+lipid solvent extraction and 2) lipid solvent extraction+filtration. Route 1 revealed to be much better since the used membrane for filtration was able to permeate the sugar aqueous phase and retained the fraction containing lipids; after this, an extraction required a much lower amount of solvent and a better recovering yield...
August 2017: Bioresource Technology
https://www.readbyqxmd.com/read/28343780/propagation-of-brazilian-zika-virus-strains-in-static-and-suspension-cultures-using-vero-and-bhk-cells
#16
Alexander Nikolay, Leda R Castilho, Udo Reichl, Yvonne Genzel
The recent spread of Zika virus (ZIKV) in the Americas and the Pacific has reached alarming levels in more than 60 countries. However, relatively little is known about the disease on a virological and epidemiological level and its consequences for humans. Accordingly, a large demand for in vitro derived Brazilian ZIKV material to support in vitro and in vivo studies has arisen. However, a prompt supply of ZIKV and ZIKV antigens cannot be guaranteed as the production of this virus typically using Vero or C6/36 cell lines remains challenging...
March 23, 2017: Vaccine
https://www.readbyqxmd.com/read/28324833/simplified-purification-approach-of-urinary-neutrophil-gelatinase-associated-lipocalin-by-tangential-flow-filtration-and-ion-exchange-chromatography
#17
Kunal K Shukla, Shamkant B Badgujar, Paresh B Bhanushali, Sushma G Sabharwal
This investigation reports a simplified approach for the purification of urinary siderocalin known as neutrophil gelatinase-associated lipocalin (NGAL). Urinary NGAL was purified by tangential flow filtration and ion exchange chromatography. Isolated NGAL was analyzed by SDS-PAGE, immunoblotting and mass spectrometry (MS). The relative molecular mass of NGAL is 23674Da. Peptide mass fingerprinting of the purified NGAL yielded peptides that partially matched with known sequence of P80188 (NGAL_HUMAN). The tryptic digestion profile of isolated NGAL infers that it may be unique and additive molecule in the dictionary of urinary proteins...
April 15, 2017: Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
https://www.readbyqxmd.com/read/28213251/on-line-monitoring-of-in-vitro-oral-bioaccessibility-tests-as-front-end-to-liquid-chromatography-for-determination-of-chlorogenic-acid-isomers-in-dietary-supplements
#18
Daniel Kremr, David J Cocovi-Solberg, Petra Bajerová, Karel Ventura, Manuel Miró
A novel fully automated in-vitro oral dissolution test assay as a front-end to liquid chromatography has been developed and validated for on-line chemical profiling and monitoring of temporal release profiles of three caffeoylquinic acid (CQA) isomers, namely, 3-CQA,4-CQA and 5-CQA, known as chlorogenic acids, in dietary supplements. Tangential-flow filtration is harnessed as a sample processing approach for on-line handling of CQA containing extracts of hard gelatin capsules and introduction of protein-free samples into the liquid chromatograph...
May 1, 2017: Talanta
https://www.readbyqxmd.com/read/28177193/manufacturing-of-recombinant-adeno-associated-viruses-using-mammalian-expression-platforms
#19
REVIEW
Marc-André Robert, Parminder S Chahal, Alexandre Audy, Amine Kamen, Rénald Gilbert, Bruno Gaillet
Manufacturing practices for recombinant adeno-associated viruses (AAV) have improved in the last decade through the development of new platforms in conjunction with better production and purification methods. In this review, we discuss the advantages and limitations of the most popular systems and methods employed with mammalian cell platforms. Methods and systems such as transient transfection, packaging and producer cells and adenovirus and herpes simplex virus are described. In terms of best production yields, they are comparable with about 10(4) -10(5) vector genomes produced per cell but transient transfection of HEK293 cells is by far the most commonly used...
March 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28159614/shear-contributions-to-cell-culture-performance-and-product-recovery-in-atf-and-tff-perfusion-systems
#20
Samantha Wang, Scott Godfrey, Janani Ravikrishnan, Henry Lin, Jens Vogel, Jon Coffman
Achievement of a robust and scalable cell retention device remains a challenge in perfusion systems. Of the two filtration systems commonly used, tangential flow filtration (TFF) systems often have an inferior product sieving profile compared to alternating tangential flow filtration (ATF) systems, which is typically attributed to the ATF's unique alternating flow. Here, we demonstrate that observed performance differences between the two systems are a function of cell lysis and not the alternating flow as previously thought...
March 20, 2017: Journal of Biotechnology
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