Vaccine Plasmid Topology Monitoring by Capillary Gel Electrophoresis.

Abstract:

BACKGROUND:Plasmid DNA has been widely used in vaccination as well as in cell and gene therapy. It exists in multiple isoforms, including supercoiled, nicked or open circular and linear forms. Regulatory agencies recommend having more than 80% of the supercoiled isoform for the bulk release of plasmid products; thus, it should be analyzed accordingly. METHODS AND RESULTS:The traditional analysis method for plasmid DNA is agarose gel electrophoresis. However, due to time-consuming manual sample loading, visualization, and data analysis, it has limitations in obtaining consistently quantitative results. In this short communication, we introduced a fast, sensitive, and robust plasmid analysis method using capillary gel electrophoresis with laser-induced fluorescence detection (CGE-LIF). CGE-LIF analysis of the supercoiled isoform and its open circular counterpart was completed in 20 minutes with excellent sensitivity by using a common fluorescent DNA binding dye. The advantage of the method was demonstrated by the purity analysis of two large plasmids (7 kb and 10 kb). The fully automated sample loading, separation and data analysis featured enhanced assay repeatability and ease of quantitation over agarose gel electrophoresis. CONCLUSION:As a worked example, analysis of plasmid samples treated at elevated temperature during an accelerated stability test also demonstrated the applicability of CGE-LIF to monitor plasmid topology and possible degradation.

journal_name

Curr Mol Med

authors

Cook KS,Luo J,Guttman A,Thompson L

doi

10.2174/1566524020666200427110452

subject

Has Abstract

pub_date

2020-04-27 00:00:00

eissn

1566-5240

issn

1875-5666

pii

CMM-EPUB-106158

pub_type

信件
  • Mining the genome for susceptibility to complex neurological disorders.

    abstract::Genome-wide association studies (GWAS) have become increasingly widely used to determine regions of the genome which may contain loci influencing the risk of neurological disorders. While linkage studies have identified genes that cause a number of Mendelian disorders, linkage analysis is less well suited for the more...

    journal_title:Current molecular medicine

    pub_type: 杂志文章,评审

    doi:10.2174/156652409789105534

    authors: Gubitz AK,Gwinn K

    更新日期:2009-09-01 00:00:00

  • Analysis of the Differential Expression Patterns of Sumoylation Enzymes E1, E2 and E3 in Ocular Cell Lines.

    abstract:PURPOSE:Protein sumoylation is a well established regulatory mechanism to control many cellular processes such as chromatin structure dynamics, transcriptional regulation of gene expression, cell proliferation and differentiation, cell transformation and carcinogenesis, autophagy and senescence. In the vertebrate visio...

    journal_title:Current molecular medicine

    pub_type: 杂志文章

    doi:10.2174/1566524019666190112143636

    authors: Nie Q,Xie J,Gong X,Luo Z,Wang L,Liu F,Xiang JW,Xiao Y,Fu JL,Liu Y,Chen Z,Yang L,Chen H,Gan Y,Li DW

    更新日期:2018-01-01 00:00:00

  • Frequency of Interleukins IL1ß/IL18 and Inflammasome NLRP1/NLRP3 Polymorphisms in Sickle Cell Anemia Patients and their Association with Severity Score.

    abstract:BACKGROUND:Interleukins IL1ß/IL18 and Inflammasome NLRP1/NLRP3 polymorphisms can change the course of multiple human diseases, both inflammatory as infectious. SNPs these proteins were associated with the constructive activation of the Inflammasome and excessive production of IL-1β induce a serious autoinflammatory dis...

    journal_title:Current molecular medicine

    pub_type: 杂志文章

    doi:10.2174/1566524019666190826143749

    authors: de Almeida E,Frantz SR,Cesar P,Tarragô AM,de Amorim Xabregas L,Garcia NP,Costa AG,de Paula EV,Malheiro A

    更新日期:2019-01-01 00:00:00

  • Overview of the molecular bases of resistance to chemotherapy in liver and gastrointestinal tumours.

    abstract::Primary malignancies of the liver and the gastrointestinal tract constitute one of the main health problems worldwide. Together, these types of tumour are the first cause of death due to cancer, followed by lung and breast cancer respectively. One important limitation in the treatment of these tumours is that, with a ...

    journal_title:Current molecular medicine

    pub_type: 杂志文章,评审

    doi:10.2174/156652409789839125

    authors: Marin JJ,Romero MR,Martinez-Becerra P,Herraez E,Briz O

    更新日期:2009-12-01 00:00:00

  • Elevated Interleukin 37 Expression Associated With Disease Activity in HLA-B27 Associated Anterior Uveitis and Idiopathic Anterior Uveitis.

    abstract:BACKGROUND & OBJECTIVE:Interleukin 37 (IL-37) is an important regulator of the anti-inflammatory T-cell response. In this study, we investigated its expression and function in peripheral blood mononuclear cells (PBMCs) of patients with HLA-B27 associated acute anterior uveitis (AAU) and idiopathic AAU. METHODS:15 pati...

    journal_title:Current molecular medicine

    pub_type: 杂志文章

    doi:10.2174/1566524018666180207152941

    authors: Zhao B,Zhang S,Chen W,Zhang R,Jiang R,Zhang R,Chen L

    更新日期:2018-03-09 00:00:00

  • Aging and Proteins: What Does Proteostasis Have to Do with Age?

    abstract::The world is aging and we must face the challenges that this brings. One of the reasons for the increasing aging of the world's population is the increase in life expectancy and, since we live longer, it is of paramount importance to live well and to prevent age-associated diseases. In this way, it is crucial to impro...

    journal_title:Current molecular medicine

    pub_type: 杂志文章,评审

    doi:10.2174/1566524018666180907162955

    authors: Magalhaes S,Goodfellow BJ,Nunes A

    更新日期:2018-01-01 00:00:00

  • Ipsilateral hippocampal proteomics reveals mitochondrial antioxidative stress impairment in cortical-lesioned chronic mild stressed rats.

    abstract::In this study, a two-dimensional gel-based proteomic approach was applied to profile the protein alterations underlying the significant adverse effects from post-stroke depression (PSD). In view of the close association between left prefrontal cortical dysfunction and PSD, a PSD rat model was constructed through a lef...

    journal_title:Current molecular medicine

    pub_type: 杂志文章

    doi:10.2174/1566524014666141021143333

    authors: Pan J,Liu H,Zhou J,Liu Z,Yang Y,Peng Y,You H,Yang D,Xie P

    更新日期:2014-01-01 00:00:00

  • Cyclic ADP-ribose (cADPR) and nicotinic acid adenine dinucleotide phosphate (NAADP): novel regulators of Ca2+-signaling and cell function.

    abstract::Ca2+ ions are involved in the regulation of many diverse functions in animal and plant cells, e.g. muscle contraction, secretion of neurotransmitters, hormones and enzymes, fertilization of oocytes, and lymphocyte activation and proliferation. The intracellular Ca2+ concentration can be increased by different molecula...

    journal_title:Current molecular medicine

    pub_type: 杂志文章,评审

    doi:10.2174/1566524024605707

    authors: Guse AH

    更新日期:2002-05-01 00:00:00

  • Targeting the tumor proteasome as a mechanism to control the synthesis and bioactivity of matrix macromolecules.

    abstract::Extracellular matrices (ECMs) are dynamic structures that provide cells not only with a structural support but, importantly, exhibit significant functional roles in the control of key cellular events such as adhesion, migration, proliferation, differentiation, and survival. In tumors, matrix effectors such as proteogl...

    journal_title:Current molecular medicine

    pub_type: 杂志文章,评审

    doi:10.2174/156652412802480943

    authors: Skandalis SS,Aletras AJ,Gialeli C,Theocharis AD,Afratis N,Tzanakakis GN,Karamanos NK

    更新日期:2012-09-01 00:00:00

  • Medullary thyroid cancer: a promising model for targeted therapy.

    abstract::In recent years, the clinical validation of molecular targeted therapies inhibiting the action of pathogenic tyrosine kinase (TK) has been one of the most exciting developments in cancer research. In this context, medullary thyroid carcinoma (MTC) represents a promising model. It is well known that in MTC, the RET rec...

    journal_title:Current molecular medicine

    pub_type: 杂志文章,评审

    doi:10.2174/156652410792630607

    authors: Torino F,Paragliola RM,Barnabei A,Corsello SM

    更新日期:2010-10-01 00:00:00

  • Apoptosis and oxidative stress-related diseases: the p66Shc connection.

    abstract::p66Shc is the only known proapoptotic member of the Shc protein family of molecular adaptors. Through its redox activity, p66Shc oxidates cytochrome-c, leading to increased ROS production and, eventually, to apoptosis. p66Shc has been implicated in the control of oxidative stress and life span in mammals. In this revi...

    journal_title:Current molecular medicine

    pub_type: 杂志文章,评审

    doi:10.2174/156652409787847254

    authors: Pellegrini M,Baldari CT

    更新日期:2009-04-01 00:00:00

  • Continuing education of the immune system--dendritic cells, immune regulation and tolerance.

    abstract::T cells, as they develop in the thymus come to express antigen receptors. The specificity of these receptors cannot be predicted and must include many with potential anti-self reactivity. Those that encounter self-antigens, in association with self-MHC (major histocompatibility complex), with high affinity are inactiv...

    journal_title:Current molecular medicine

    pub_type: 杂志文章,评审

    doi:10.2174/1566524013363573

    authors: Huang FP,MacPherson GG

    更新日期:2001-09-01 00:00:00

  • Molecular mistletoe therapy: friend or foe in established anti-tumor protocols? A multicenter, controlled, retrospective pharmaco-epidemiological study in pancreas cancer.

    abstract::Mistletoe is often used as complementary therapy in oncology. The anti-tumor effects of mistletoe (Iscador) are well documented in-vitro in respect to inhibition of cell proliferation, induction of apoptosis, segmental activation of immune competent cells and trapping of chemotherapeutic drugs within cancer cells by m...

    journal_title:Current molecular medicine

    pub_type: 杂志文章,多中心研究

    doi:10.2174/156652410791317057

    authors: Matthes H,Friedel WE,Bock PR,Zänker KS

    更新日期:2010-06-01 00:00:00

  • High Glucose Affects the Cytotoxic Potential of Rapamycin, Metformin and Hydrogen Peroxide in Cultured Human Mesenchymal Stem Cells.

    abstract:BACKGROUND:Oxidative stress and chronic hyperglycemia are two major side effects of type 2 diabetes affecting all cell types including mesenchymal stem cells (MSCs). As a cell therapy choice, understanding the behavior of MSCs will provide crucial information for efficient treatment. METHODS:Placental mesenchymal stem...

    journal_title:Current molecular medicine

    pub_type: 杂志文章

    doi:10.2174/1566524019666190722115842

    authors: Roohi A,Nikougoftar M,Montazeri H,Navabi S,Shokri F,Ostad SN,Ghahremani MH

    更新日期:2019-01-01 00:00:00

  • Adipokine Visfatin's Role in Pathogenesis of Diabesity and Related Metabolic Derangements.

    abstract::Visfatin is one of the prominent adipokines secreted by adipose tissue. The level of visfatin increases significantly in persons with obesity owing to increased body mass index (BMI). During obesity, the adipocytes, which populate adipose tissue, undergo hypertrophy and hyperplasia and secrete a number of adipocytokin...

    journal_title:Current molecular medicine

    pub_type: 杂志文章,评审

    doi:10.2174/1566524018666180705114131

    authors: Kumari B,Yadav UCS

    更新日期:2018-01-01 00:00:00

  • Expression of Dominant Negative K6W-Ubiquitin in the Lens Epithelium via an Adenoviral Vector Delays Posterior Capsule Opacification in a Rabbit Model.

    abstract:PURPOSE:Ubiquitin is involved in cell proliferation and differentiation, and the objective of this study is to investigate the potential of dominant negative Ubiquitin (Ub) with a lysine to tryptophan mutation at the 6 position (K6W) through an adenoviral expression vector in the prevention of posterior capsule opacifi...

    journal_title:Current molecular medicine

    pub_type: 杂志文章

    doi:10.2174/1566524017666170331163751

    authors: Bao X,Hou M,Peng R,Luo F,Wu M

    更新日期:2017-01-01 00:00:00

  • Early onset gastric cancer: on the road to unraveling gastric carcinogenesis.

    abstract::Gastric cancer is thought to result from a combination of environmental factors and the accumulation of specific genetic alterations due to increasing genetic instability, and consequently affects mainly older patients. Less than 10% of patients present with the disease before 45 years of age (early onset gastric carc...

    journal_title:Current molecular medicine

    pub_type: 杂志文章,评审

    doi:10.2174/156652407779940503

    authors: Milne AN,Sitarz R,Carvalho R,Carneiro F,Offerhaus GJ

    更新日期:2007-02-01 00:00:00

  • Studying Adsorption and Cellular Toxicity of Boron Nitride Nanostructure versus Melphalan Anti-ovarian Cancer Drug.

    abstract:BACKGROUND:Inclusion of anticancer drugs into biocompatible nanoparticulate carriers decreases the general toxicity and improves the efficacy of clinical treatments due to the reduction of soluble circulating free drug. METHODS:In addition, removal of emerging drug contaminants from wastewaters is a necessity that sho...

    journal_title:Current molecular medicine

    pub_type: 杂志文章

    doi:10.2174/1566524021666210111104428

    authors: Fu M,Tayebee R,Saberi S,Nourbakhsh N,Esmaeili E,Maleki B,Vatanpour HR

    更新日期:2021-01-10 00:00:00

  • p53 peptide prevents LITAF-induced TNF-alpha-mediated mouse lung lesions and endotoxic shock.

    abstract::Abnormal and prolonged inflammatory reaction is seen in a wide variety of disorders, and high level of Tumor Necrosis Factor alpha (TNF-α) has been linked to these disorders. Therefore, modulation of TNF-α expression is important in the regulation of inflammatory disorders. In our previous study, we have shown that a ...

    journal_title:Current molecular medicine

    pub_type: 杂志文章

    doi:10.2174/156652411796268731

    authors: Tang X,O'Reilly A,Asano M,Merrill JC,Yokoyama KK,Amar S

    更新日期:2011-08-01 00:00:00

  • Endogenous Factors in the Recovery of Reproductive Function After Testicular Injury and Cancer.

    abstract::The testes are one of the most delicate organs in the male body and highly susceptible to the exogenous influences capable of inducing cell damage. Cancer therapies are well known to negatively affect the male reproductive tract with a severe impairment of spermatogenesis and infertility. The present work aimed to sys...

    journal_title:Current molecular medicine

    pub_type: 杂志文章,评审

    doi:10.2174/1566524016666160802150400

    authors: Silva AM,Correia S,Socorro S,Maia CJ

    更新日期:2016-01-01 00:00:00

  • Association of Genetic Variants with Colorectal Cancer in the Extended MENA Region: A Systematic Review.

    abstract:BACKGROUND:Colorectal cancer (CRC) is the third most common cancer worldwide and the third leading cause of cancer-related death. It is a heterogeneous disease that develops through different genetic and epigenetic mechanisms. To date, no comprehensive systematic review investigating genetic risk factors for familial a...

    journal_title:Current molecular medicine

    pub_type: 杂志文章

    doi:10.2174/1566524019666191014170136

    authors: Sidenna M,Bux R,Fadl T,Ozbek U,Zayed H

    更新日期:2020-01-01 00:00:00

  • Rapid differentiation of chondroitin sulfate isomers by gas-phase hydrogen/deuterium exchange.

    abstract::Chondroitin sulfate (CS)-glycosaminoglycans (GAGs) are linear, negatively charged polysaccharides attached to CS proteoglycans that make up a major component of biological matrices throughout both central and peripheral tissues. The position of their attached sulfate groups to the CS disaccharide is predicted to influ...

    journal_title:Current molecular medicine

    pub_type: 杂志文章

    doi:10.2174/1566524020666200915110707

    authors: Alonge K,Harkewicz R,Guttman M

    更新日期:2020-09-15 00:00:00

  • Distinct and Overlapping Roles for E2F Family Members in Transcription, Proliferation and Apoptosis.

    abstract::Since the discovery almost fifteen years ago that E2F transcription factors are key targets of the retinoblastoma protein (RB), studies of the E2F family have uncovered critical roles in the control of transcription, cell cycle and apoptosis. E2F proteins are encoded by at least eight genes, E2F1 through E2F8. While s...

    journal_title:Current molecular medicine

    pub_type: 杂志文章,评审

    doi:10.2174/1566524010606070739

    authors: DeGregori J,Johnson DG

    更新日期:2006-11-01 00:00:00

  • The role of transcriptome analysis in pre-clinical toxicology.

    abstract::A major benefit of the genomics revolution in biomedical research has been the establishment of transcriptome analysis as an enabling technology in the drug development process. Nowhere in the realm of drug development has the expectation of the impact of transcriptome analysis been greater than in the area of pre-cli...

    journal_title:Current molecular medicine

    pub_type: 杂志文章,评审

    doi:10.2174/1566524053152825

    authors: Searfoss GH,Ryan TP,Jolly RA

    更新日期:2005-02-01 00:00:00

  • Mesenchymal stem cells regulate cytoskeletal dynamics and promote cancer cell invasion through low dose nitric oxide.

    abstract::Bone marrow-derived mesenchymal stem cells (BMSCs) can be recruited to tumor sites and integrate into the stroma of tumors. When co-cultured with BMSCs, otherwise weakly metastatic nasopharyngeal carcinoma cells (NPC) showed improved metastatic ability. BMSCs in the tumor environment displayed the characteristics of m...

    journal_title:Current molecular medicine

    pub_type: 杂志文章

    doi:10.2174/1566524014666140724102301

    authors: Zhang L,Sun J,Liu Z,Dai Y,Luo Z,Jiang X,Li Z,Li Y,Cao P,Zhou Y,Zeng Z,Tang A,Li X,Li X,Xiang J,Li G

    更新日期:2014-01-01 00:00:00

  • Forward programming of pluripotent stem cells to neurons.

    abstract::Pluripotent stem cells (PSCs) are powerful tools for studying developmental biology and neuronal diseases. Conventional differentiation protocols require several intermediate states and different culture conditions, inefficiently generating mixed subtypes of neuronal cells with immature characteristics. Direct program...

    journal_title:Current molecular medicine

    pub_type: 杂志文章

    doi:10.2174/1566524020666200421115251

    authors: Gu J,Cromer B,Sumer H

    更新日期:2020-04-21 00:00:00

  • New Horizons in the Etiopathogenesis and Non-Invasive Diagnosis of Endometriosis.

    abstract::Endometriosis is one of the most common gynecological inflammatory diseases, occurring in adolescents and women in the reproductive age group and leading to infertility. The precise etiopathogenesis of endometriosis is unknown, but several theories concerning the phenomena involved in its development have been propose...

    journal_title:Current molecular medicine

    pub_type: 杂志文章,评审

    doi:10.2174/1566524015666150921105218

    authors: Zubrzycka A,Zubrzycki M,Janecka A,Zubrzycka M

    更新日期:2015-01-01 00:00:00

  • Von Hippel-Lindau disease.

    abstract::Germline mutations in the VHL tumour suppressor gene may cause a variety of phenotypes including von Hippel-Lindau (VHL) disease, familial phaeochromocytoma and inherited polycythaemia. VHL disease is a multisystem familial cancer syndrome and is the commonest cause of familial renal cell carcinoma (RCC). VHL disease ...

    journal_title:Current molecular medicine

    pub_type: 杂志文章,评审

    doi:10.2174/1566524043359827

    authors: Maher ER

    更新日期:2004-12-01 00:00:00

  • Past, present and future strategies of immunotherapy in gynecological malignancies.

    abstract::Recently, the combining of different drugs has greatly improved response and survival rates in gynecological malignancies. Results are however far from being satisfactory. Treatments used in case of advanced or recurrent disease offer limited results in terms of long-term responses. The urgent need for new and more ef...

    journal_title:Current molecular medicine

    pub_type: 杂志文章

    doi:10.2174/1566524011313040014

    authors: Bellati F,Napoletano C,Ruscito I,Visconti V,Antonilli M,Gasparri ML,Zizzari IG,Rahimi H,Palaia I,Rughetti A,Benedetti Panici P,Nuti M

    更新日期:2013-05-01 00:00:00

  • Role of innate immunity in triggering and tuning of autoimmune diabetes.

    abstract::Type 1 diabetes (T1D) is an organ-specific autoimmune disease resulting from the specific destruction of insulin-producing pancreatic beta-cells, culminating in a state of hypoinsulinemia and hyperglycemia. Pathogenesis of T1D comprises complex series of events from the initial sensitization of antigen-presenting cell...

    journal_title:Current molecular medicine

    pub_type: 杂志文章,评审

    doi:10.2174/156652409787314471

    authors: Kim HS,Lee MS

    更新日期:2009-02-01 00:00:00