Abstract:
:Engineered human T-cells are a promising therapeutic modality for cancer immunotherapy. T-cells expressing chimeric antigen receptors combined with additional genes to enhance T-cell proliferation, survival, or tumor targeting may further improve efficacy but require multiple stable gene transfer events. Methods are therefore needed to increase production efficiency for multiplexed engineered cells. In this work, we demonstrate multiplexed, non-viral gene transfer to a human T-cell line with efficient selection (∼ 50%) of cells expressing up to three recombinant open reading frames. The efficient introduction of multiple genes to T-cells was achieved using the Sleeping Beauty transposon system delivered in minicircles by nucleofection. We demonstrate rapid selection for engineered cells using methotrexate (MTX) and a mutant human dihydrofolate reductase resistant to methotrexate-induced metabolic inhibition. Preferential amplification of cells expressing multiple transgenes was achieved by two successive rounds of increasing MTX concentration. This non-viral gene transfer method with MTX step selection can potentially be used in the generation of clinical-grade T-cells housing multiplexed genetic modifications.
journal_name
Biotechnol Bioengjournal_title
Biotechnology and bioengineeringauthors
Kacherovsky N,Liu GW,Jensen MC,Pun SHdoi
10.1002/bit.25538subject
Has Abstractpub_date
2015-07-01 00:00:00pages
1429-36issue
7eissn
0006-3592issn
1097-0290journal_volume
112pub_type
杂志文章abstract::The progression of liver disease from normal to cirrhotic state is characterized by modulation of the stiffness of the extracellular matrix (ECM). Mimicking this modulation in vitro scaffold could provide a better insight into hepatic cell behavior. In this study, interpenetrating poly(N-isopropylacrylamide-co-gelatin...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.27218
更新日期:2020-02-01 00:00:00
abstract::Animal cell culture is widely used in biotechnology for the production of many biological products. In situ microscopes acquire images directly from cell suspensions and analyze the images in matters of cell concentration, cell size distribution and cell morphology. Their applicability was already proven for yeast and...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.21523
更新日期:2008-01-01 00:00:00
abstract::In the process of producing ethanol from lignocellulosic materials such as wheat straw, compounds that can act inhibitory to enzymatic hydrolysis and to cellular growth may be generated during the pretreatment. Ethanol production was evaluated on pretreated wheat straw hydrolysate using four different recombinant Sacc...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.21100
更新日期:2007-02-01 00:00:00
abstract::Incremental truncation is a method for constructing libraries of every one base pair truncation of a segment of DNA. Incremental truncation libraries can be created using a time-dependent nuclease method or through the incorporation of alpha-phosphothioate dNTPs by PCR or by primer extension (THIO(pcr) truncation and ...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.10604
更新日期:2003-06-05 00:00:00
abstract::Reduction and oxidation of steroids in the human gut are catalyzed by hydroxysteroid dehydrogenases of microorganisms. For the production of 12-ketochenodeoxycholic acid (12-Keto-CDCA) from cholic acid the biocatalytic application of the 12α-hydroxysteroid dehydrogenase of Clostridium group P, strain C 48-50 (HSDH) is...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.23047
更新日期:2011-06-01 00:00:00
abstract::This paper proposes a new metabolic model for acetate uptake by a mixed culture of phosphate- and glycogen-accumulating organisms (PAOs and GAOs) under anaerobic conditions. The model uses variable overall stoichiometry based on the assumption that PAOs may have the ability of using the glyoxylate pathway to produce t...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.10765
更新日期:2003-11-05 00:00:00
abstract::A flow cytometric kinetic study of hybridoma growth and monoclonal antibody production is presented, along with the influence of glutamine on intracellular responses such as (relative) cell size, and cell RNA and total protein content. Specific findings are: (1) RNA content remained constant throughout the growth phas...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.260360109
更新日期:1990-06-05 00:00:00
abstract::A simple approach was developed to determine the half-saturation coefficient for dissolved oxygen (K(DO)) for three bacteria by maintaining a constant oxygen concentration in continuous culture, and employing a dynamic method to obtain the specific growth rate (mu) for each species. Measurement of mu at selected disso...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.260250209
更新日期:1983-02-01 00:00:00
abstract::This paper describes batch and semicontinuous acetic acid fermentations for wine vinegar production carried out with Acetobacter pasteurianus, and an industrial strain using a noncommercial 100-L bubble column reactor equipped with a novel type of gas-liquid dynamic sparger. Results showed acetification rates with thi...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/(sici)1097-0290(19990420)63:2<141::aid-bit
更新日期:1999-04-20 00:00:00
abstract::High throughput pretreatment (HTPH) and enzymatic hydrolysis systems are now vital for screening large numbers of biomass samples to investigate biomass recalcitrance over various pretreatment and enzymatic hydrolysis conditions. Although hydrothermal pretreatment is currently being employed in most high throughput ap...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.24951
更新日期:2013-11-01 00:00:00
abstract::A soil with aged contamination from lubricating oil (LO) and polychlorinated biphenyls (PCBs) was treated in a bioslurry reactor to investigate in-soil biosurfactant production by Pseudomonas aeruginosa, the most abundant indigenous, culturable, hydrocarbon-degrading microorganism. After 2 days of growth on LO, a depl...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.20295
更新日期:2004-12-30 00:00:00
abstract::Macroencapsulation technology has been an attractive topic in the field of treatment for Type 1 diabetes due to mechanical stability, versatility, and retrievability of the macro-capsule design. Macro-capsules can be categorized into extravascular and intravascular devices, in which solute transport relies either on d...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章,评审
doi:10.1002/bit.25895
更新日期:2016-07-01 00:00:00
abstract::Beta defensins are antimicrobial peptides (AMPs) with a broad spectrum antimicrobial behavior against pathogens while having minimal tendency to incur pathogen resistance. Human β-defensin 28 (hBD28) is a strongly cationic AMP and hence hypothesized to be highly effective in permeabilizing negatively-charged pathogen ...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.22970
更新日期:2011-03-01 00:00:00
abstract::The application of the unscented Kalman filter to control starvation-induced programmed cell death-apoptosis-in Chinese hamster ovary cells was investigated. Neural network-based sensitivity analysis identified glutamine and asparagine as two major amino acids that play a key role in the suppression of apoptosis. Dyna...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.10250
更新日期:2002-06-20 00:00:00
abstract::In this article, we consider the impact on downstream process design resulting from the use of metabolically engineered yeast strains. We address the issue of how manipulation of cell wall permeability can improve the release and subsequent recovery of heterologous products produced in yeast. ...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章,评审
doi:
更新日期:1998-04-20 00:00:00
abstract::The feasibility of anaerobic methanethiol (MT) degradation at elevated sodium concentrations was investigated in a mesophilic (30 degrees C) lab-scale upflow anaerobic sludge bed (UASB) reactor, inoculated with estuarine sediment originating from the Wadden Sea (The Netherlands). MT was almost completely degraded (>95...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.21367
更新日期:2007-09-01 00:00:00
abstract::In biotechnology and biomedicine reliable models of cell proliferation kinetics need to capture the relevant phenomena taking place during the mitotic cycle. To this aim, a novel mathematical model helpful to investigate the intrinsic kinetics of in vitro culture of adherent cells up to confluence is proposed in this ...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.24351
更新日期:2012-03-01 00:00:00
abstract::Being biosurfactants, rhamnolipids create severe foaming when produced in aerobic Pseudomonas aeruginosa fermentation. The necessary reduction of aeration causes oxygen limitation and restricts cell and product concentrations. In this study, we evaluate the new strategy of rhamnolipid production under denitrification ...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/1097-0290(20010105)72:1<25::aid-bit4>3.0.c
更新日期:2001-01-05 00:00:00
abstract::D-Methionine was converted to L-methionine in a reaction system where four enzymes were used. D-amino acid oxidase (D-AAO) from Arthrobacter protophormiae was used for the complete conversion of D-methionine to 2-oxo-4-methylthiobutyric acid. Catalase was added to prevent 2-oxo-4-methylthiobutyric acid decarboxylation...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.21501
更新日期:2007-12-01 00:00:00
abstract::In this study, we assessed the importance of cytoskeleton organization in the mammalian cells used to produce therapeutic proteins. Two cytoskeletal genes, Actin alpha cardiac muscle 1 (ACTC1) and a guanosine triphosphate GTPase-activating protein (TAGAP), were found to be upregulated in highly productive therapeutic ...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.27277
更新日期:2020-04-01 00:00:00
abstract::We consider the application of high-throughput experimentation (HTE), including combinatorial methods, to catalyst discovery and early-phase optimization. While combinatorial- and parallel-testing methods have an already substantial history in catalysis, recent work by several groups promises significant efficiency ga...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/(sici)1097-0290(1998)61:4<203::aid-cc3>3.0
更新日期:1998-01-01 00:00:00
abstract::An extensive investigation of anaerobic methane fermentation requires identifying the relationship between the physical environment and biological process. In this study, a computational fluid dynamics (CFD) technique was used to characterize bacterial fermentation mechanisms intertwined with mixing and heat transfer ...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.24551
更新日期:2012-11-01 00:00:00
abstract::The implementation of control algorithms to batch bioreactors is often complicated by variations in process dynamics that occur during the course of fermentation. Such a wide operating range often renders the performance of fixed gain proportional-integral-differential (PID) controllers unsatisfactory. In this work, d...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.260320415
更新日期:1988-08-05 00:00:00
abstract::Mevalonate is a useful metabolite synthesized from three molecules of acetyl-CoA, consuming two molecules of NADPH. Escherichia coli ( E. coli) catabolizes glucose to acetyl-CoA via several routes, such as the Embden-Meyerhof-Parnas (EMP) and the oxidative pentose phosphate (oxPP) pathways. Although the oxPP pathway s...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.26923
更新日期:2019-05-01 00:00:00
abstract::Two metabolic engineering tools, namely gene inactivation and gene overexpression, were employed to examine the effects of two genetic modifications on the fermentation characteristics of Clostridium acetobutylicum. Inactivation of the butyrate kinase gene (buk) was examined using strain PJC4BK, while the combined eff...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:
更新日期:2000-01-05 00:00:00
abstract::A technology has been developed to achieve optimal attachment of adhesion-independent lymphocytes to microcarrier beads. The activation of T-lymphocytes by concanavalin A was tested under microgravity conditions in an experiment carried out in space during the first Spacelab Life Science Mission. Activation, measure...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.260400815
更新日期:1992-10-20 00:00:00
abstract::The transient and steady-state responses of hybridoma growth and metabolism to glutamine pulse and step changes have been examined. Metabolic quotients are reported for oxygen, glucose, lactate, ammonia, glutamine, alanine, and other amino acids. The specific glutamine consumption rate increased rapidly after all glut...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.260330414
更新日期:1989-01-20 00:00:00
abstract::A number of bioreactor configurations have been developed for the manufacture of products from mammalian cell hosts. Even in the most efficient of these, however, problems such as nutrient exhaustion, growth factor deprivation, and toxin accumulations may arise. Consequently, the current effort focused on the feasibil...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/(sici)1097-0290(20000305)67:5<555::aid-bit
更新日期:2000-03-05 00:00:00
abstract::Mucopolysaccharidosis (MPS) Type II is caused by mutations in the gene encoding the lysosomal enzyme, iduronate 2-sulfatase (IDS). The majority of MPSII cases affect the brain. However, enzyme replacement therapy with recombinant IDS does not treat the brain, because IDS is a large molecule drug that does not cross th...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.25289
更新日期:2014-11-01 00:00:00
abstract::Chlorella sp. strain VJ79 was isolated from a total heterotrophic count of a wastewater collector. It grows autotrophically, heterotrophically, and mixotrophically on a variety of organic substrates. Glucose and serine promote a mixotrophic growth from which the yield is higher than the sum of autotrophic and heterotr...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.260260707
更新日期:1984-07-01 00:00:00