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 Medjournal_title
Current molecular medicineauthors
Cook KS,Luo J,Guttman A,Thompson Ldoi
10.2174/1566524020666200427110452subject
Has Abstractpub_date
2020-04-27 00:00:00eissn
1566-5240issn
1875-5666pii
CMM-EPUB-106158pub_type
信件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
更新日期:2009-09-01 00:00:00
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
更新日期:2018-01-01 00:00:00
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
更新日期:2019-01-01 00:00:00
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
更新日期:2009-12-01 00:00:00
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
更新日期:2018-03-09 00:00:00
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
更新日期:2018-01-01 00:00:00
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
更新日期:2014-01-01 00:00:00
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
更新日期:2002-05-01 00:00:00
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
更新日期:2012-09-01 00:00:00
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
更新日期:2010-10-01 00:00:00
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
更新日期:2009-04-01 00:00:00
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
更新日期:2001-09-01 00:00:00
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
更新日期:2010-06-01 00:00:00
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
更新日期:2019-01-01 00:00:00
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
更新日期:2018-01-01 00:00:00
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
更新日期:2017-01-01 00:00:00
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
更新日期:2007-02-01 00:00:00
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
更新日期:2021-01-10 00:00:00
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
更新日期:2011-08-01 00:00:00
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
更新日期:2016-01-01 00:00:00
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
更新日期:2020-01-01 00:00:00
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
更新日期:2020-09-15 00:00:00
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
更新日期:2006-11-01 00:00:00
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
更新日期:2005-02-01 00:00:00
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
更新日期:2014-01-01 00:00:00
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
更新日期:2020-04-21 00:00:00
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
更新日期:2015-01-01 00:00:00
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
更新日期:2004-12-01 00:00:00
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
更新日期:2013-05-01 00:00:00
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
更新日期:2009-02-01 00:00:00