AI Article Synopsis
Article Abstract
pUDK-HGF, recombinant plasmid DNA encoding human HGF (hepatocyte growth factor), is a potential agent for gene therapy of ischaemic disease. Production of pUDK-HGF is essential for its clinical application. In the present paper, a large-scale manufacturing process was developed, including fermentation, cell harvest, alkaline lysis, capturing plasmid DNA with Q-Sepharose XL chromatography, size-exclusion chromatography on a Sephacryl S1000 column and refining with Source 15Q anion-exchange chromatography. The quality criteria of pUDK-HGF such as purity, concentration, homogeneity, residual RNA, chromosomal DNA, contaminated protein, endotoxin and HGF expression efficacy all were analysed and met the requirements for pharmaceutical-grade plasmid DNA.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1042/BA20060251 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Macrophage-specific in vivo RNA editing promotes phagocytosis and antitumor immunity in mice.
Sci Transl Med
January 2025
College of Pharmaceutical Sciences, State Key Laboratory of Advanced Drug Delivery and Release Systems, Zhejiang University, Hangzhou 310058, China.
Macrophages play a central role in antitumor immunity, making them an attractive target for gene therapy strategies. However, macrophages are difficult to transfect because of nucleic acid sensors that can trigger the degradation of foreign plasmid DNA. Here, we developed a macrophage-specific editing (MAGE) system by which compact plasmid DNA encoding a CasRx editor can be delivered to macrophages by a poly(β-amino ester) (PBAE) carrier to bypass the DNA sensor and enable RNA editing in vitro and in vivo.
View Article and Find Full Text PDFThe formation and architecture of surface-initiated polymer brush gene delivery complexes.
J Colloid Interface Sci
December 2024
School of Engineering and Materials Science, Queen Mary University of London, Mile End Road, London E1 4NS, United Kingdom. Electronic address:
Understanding the architecture and mechanism of assembly of polyelectrolyte-nucleic acid complexes is critical to the rational design of their performance for gene delivery. Surface-initiated polymer brushes were recently found to be particularly effective at delivering oligonucleotides and maintaining high knock down efficiencies for prolonged periods of time, in highly proliferative cells. However, what distinguishes their binding capacity for oligonucleotides from that of larger therapeutic macromolecules remains unknown.
View Article and Find Full Text PDFLong-term blood glucose control via glucose-activated transcriptional regulation of insulin analogue in type 1 diabetes mice.
Diabetes Obes Metab
January 2025
National Engineering Research Center for Biomaterials, College of Biomedical Engineering, Sichuan University, Chengdu, Sichuan, People's Republic of China.
Aim: To achieve glucose-activated transcriptional regulation of insulin analogue in skeletal muscle of T1D mice, thereby controlling blood glucose levels and preventing or mitigating diabetes-related complications.
Materials And Methods: We developed the GANIT (Glucose-Activated NFAT-regulated INSA-F Transcription) system, an innovative platform building upon the previously established intramuscular plasmid DNA (pDNA) delivery and expression system. In the GANIT system, skeletal muscle cells are genetically engineered to endogenously produce the insulin analogue INSA-F (Insulin Aspart with Furin cleavage sites).
Metal-Coordinated Histidine-Functionalized Redox-Responsive Polyethyleneimine as a Smart Gene Delivery Vector.
Mol Biotechnol
January 2025
Noncommunicable Disease Research Center, Jahrom University of Medical Sciences, Jahrom, Iran.
Despite significant advancements in gene delivery and CRISPR technology, several challenges remain. Chief among these are overcoming serum inhibition and achieving high transfection efficiency with minimal cytotoxicity. To address these issues, there is a need for novel vectors that exhibit lower toxicity, maintain stability in serum-rich environments, and effectively deliver plasmids of various sizes across diverse cell types.
View Article and Find Full Text PDFPlasmid Stability Analysis with Open-Source Droplet Microfluidics.
J Vis Exp
December 2024
Institute for Biological and Medical Engineering, Pontificia Universidad Católica de Chile;
Plasmids play a vital role in synthetic biology by enabling the introduction and expression of foreign genes in various organisms, thereby facilitating the construction of biological circuits and pathways within and between cell populations. For many applications, maintaining functional plasmids without antibiotic selection is critical. This study introduces an open-hardware-based microfluidic workflow for analyzing plasmid retention by culturing single cells in gel microdroplets and quantifying microcolonies using fluorescence microscopy.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!