Multiplexed gene transfer to a human T-cell line by combining Sleeping Beauty transposon system with methotrexate selection.

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 Bioeng

authors

Kacherovsky N,Liu GW,Jensen MC,Pun SH

doi

10.1002/bit.25538

subject

Has Abstract

pub_date

2015-07-01 00:00:00

pages

1429-36

issue

7

eissn

0006-3592

issn

1097-0290

journal_volume

112

pub_type

杂志文章
  • Gelatin interpenetration in poly N-isopropylacrylamide network reduces the compressive modulus of the scaffold: A property employed to mimic hepatic matrix stiffness.

    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

    authors: Sarkar J,Kamble SC,Patil R,Kumar A,Gosavi SW

    更新日期:2020-02-01 00:00:00

  • Online monitoring of microcarrier based fibroblast cultivations with in situ microscopy.

    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

    authors: Rudolph G,Lindner P,Gierse A,Bluma A,Martinez G,Hitzmann B,Scheper T

    更新日期:2008-01-01 00:00:00

  • Effect of compounds released during pretreatment of wheat straw on microbial growth and enzymatic hydrolysis rates.

    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

    authors: Panagiotou G,Olsson L

    更新日期:2007-02-01 00:00:00

  • Theoretical distribution of truncation lengths in incremental truncation libraries.

    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

    authors: Ostermeier M

    更新日期:2003-06-05 00:00:00

  • Biocatalytic process optimization based on mechanistic modeling of cholic acid oxidation with cofactor regeneration.

    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

    authors: Braun M,Link H,Liu L,Schmid RD,Weuster-Botz D

    更新日期:2011-06-01 00:00:00

  • Metabolic model for acetate uptake by a mixed culture of phosphate- and glycogen-accumulating organisms under anaerobic conditions.

    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

    authors: Yagci N,Artan N,Cokgör EU,Randall CW,Orhon D

    更新日期:2003-11-05 00:00:00

  • A flow-cytometric analysis of hybridoma growth and monoclonal antibody production.

    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

    authors: Dalili M,Ollis DF

    更新日期:1990-06-05 00:00:00

  • The half-saturation coefficient for dissolved oxygen: A dynamic method for its determination and its effect on dual species competition.

    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

    authors: Hao OJ,Richard MG,Jenkins D,Blanch HW

    更新日期:1983-02-01 00:00:00

  • Wine vinegar production using a noncommercial 100-litre bubble column reactor equipped with a novel type of dynamic sparger

    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

    authors: Fregapane G,Rubio-Fernandez H,Nieto J,Salvador MD

    更新日期:1999-04-20 00:00:00

  • Application of high throughput pretreatment and co-hydrolysis system to thermochemical pretreatment. Part 2: Dilute alkali.

    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

    authors: Li H,Gao X,Demartini JD,Kumar R,Wyman CE

    更新日期:2013-11-01 00:00:00

  • Stimulating in-soil rhamnolipid production in a bioslurry reactor by limiting nitrogen.

    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

    authors: Hudak AJ,Cassidy DP

    更新日期:2004-12-30 00:00:00

  • Progress and challenges in macroencapsulation approaches for type 1 diabetes (T1D) treatment: Cells, biomaterials, and devices.

    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

    authors: Song S,Roy S

    更新日期:2016-07-01 00:00:00

  • A new bioproduction route for a novel antimicrobial peptide.

    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

    authors: Tay DK,Rajagopalan G,Li X,Chen Y,Lua LH,Leong SS

    更新日期:2011-03-01 00:00:00

  • Control of starvation-induced apoptosis in Chinese hamster ovary cell cultures.

    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

    authors: Simon L,Karim MN

    更新日期:2002-06-20 00:00:00

  • Yeast cell permeabilizing beta-1,3-glucanases: A tool for the integration of downstream processes and metabolic engineering applications to yeast.

    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:

    authors: Ferrer P,Diers I,Asenjo JA,Andrews BA

    更新日期:1998-04-20 00:00:00

  • Anaerobic methanethiol degradation in upflow anaerobic sludge bed reactors at high salinity (> or =0.5 M Na(+)).

    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

    authors: van Leerdam RC,de Bok FA,Lens PN,Stams AJ,Janssen AJ

    更新日期:2007-09-01 00:00:00

  • A novel population balance model to investigate the kinetics of in vitro cell proliferation: part I. Model development.

    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

    authors: Fadda S,Cincotti A,Cao G

    更新日期:2012-03-01 00:00:00

  • Rhamnolipid production by Pseudomonas aeruginosa under denitrification: effects of limiting nutrients and carbon substrates.

    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

    authors: Chayabutra C,Wu J,Ju LK

    更新日期:2001-01-05 00:00:00

  • Biotransformation of D-methionine into L-methionine in the cascade of four enzymes.

    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

    authors: Findrik Z,Vasić-Racki D

    更新日期:2007-12-01 00:00:00

  • Influence of cytoskeleton organization on recombinant protein expression by CHO cells.

    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

    authors: Pourcel L,Buron F,Arib G,Le Fourn V,Regamey A,Bodenmann I,Girod PA,Mermod N

    更新日期:2020-04-01 00:00:00

  • Combinatorial approaches as a component of high-throughput experimentation (HTE) in catalysis research.

    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

    authors: Newsam JM,Schüth F

    更新日期:1998-01-01 00:00:00

  • Integration of mixing, heat transfer, and biochemical reaction kinetics in anaerobic methane fermentation.

    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

    authors: Wu B

    更新日期:2012-11-01 00:00:00

  • The design of controllers for batch bioreactors.

    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

    authors: Cardello RJ,San KY

    更新日期:1988-08-05 00:00:00

  • Effect of precise control of flux ratio between the glycolytic pathways on mevalonate production in Escherichia coli.

    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

    authors: Kamata K,Toya Y,Shimizu H

    更新日期:2019-05-01 00:00:00

  • Characterization of recombinant strains of the Clostridium acetobutylicum butyrate kinase inactivation mutant: need for new phenomenological models for solventogenesis and butanol inhibition?

    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:

    authors: Harris LM,Desai RP,Welker NE,Papoutsakis ET

    更新日期:2000-01-05 00:00:00

  • Activation of microcarrier-attached lymphocytes in microgravity.

    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

    authors: Bechler B,Cogoli A,Cogoli-Greuter M,Müller O,Hunzinger E,Criswell SB

    更新日期:1992-10-20 00:00:00

  • The transient responses of hybridoma cells to nutrient additions in continuous culture: II. Glutamine pulse and step changes.

    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

    authors: Miller WM,Wilke CR,Blanch HW

    更新日期:1989-01-20 00:00:00

  • Part II. Overexpression of bcl-2 family members enhances survival of mammalian cells in response to various culture insults.

    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

    authors: Mastrangelo AJ,Hardwick JM,Zou S,Betenbaugh MJ

    更新日期:2000-03-05 00:00:00

  • Insulin receptor antibody-iduronate 2-sulfatase fusion protein: pharmacokinetics, anti-drug antibody, and safety pharmacology in Rhesus monkeys.

    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

    authors: Boado RJ,Ka-Wai Hui E,Zhiqiang Lu J,Pardridge WM

    更新日期:2014-11-01 00:00:00

  • Utilization of light for the assimilation of organic matter in Chlorella sp. VJ79.

    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

    authors: Lalucat J,Imperial J,Parés R

    更新日期:1984-07-01 00:00:00