In plasmid purification methods, especially in large quantities, almost all of them include an ion exchange chromatography purification step, and the efficiency of the chromatographic purification determines the efficiency of the entire purification step. The plasmid DNA purified by traditional ion exchange chromatography has a higher endotoxin content and endotoxin can cause side effects such as inflammation or necrosis. Therefore, in vivo gene transfection requires plasmid DNA with extremely low endotoxin content. In order to reduce the endotoxin content, the plasmid DNA enriched by ion exchange chromatography needs further processing. Generally, methods such as exclusion chromatography, isopropanol precipitation, ammonium acetate precipitation, and polyethylene glycol precipitation are used to purify plasmids, but these methods are time-consuming and complicated, and the endotoxin content in the resulting plasmid DNA is still high. Therefore, Creative BioMart uses non-ionic surfactants Triton X-114 or Triton X-100 to pre-incubate the bacterial cell clarification solution at 4°C, and then purify the plasmid DNA with Fractogel EMD TMAE (M) strong anion exchange chromatography medium, after loading the column with Triton-free buffer, the endotoxin content in the plasmid DNA was reduced, and Tritonon would not remain in the purified plasmid DNA sample. This method achieves the purpose of one-step purification of plasmid DNA by anion exchange chromatography.
This method involves the separation of aqueous surfactant solution into micelle-rich and micelle-poor regions through excluded-volume interactions. External agent notably Triton X-114 is needed to maintain the inherent biological activity of the protein while reducing endotoxin level by 100-fold.
Endotoxin removal is one of the most difficult tasks downstream in protein purification. At Creative BioMart, our experienced scientists will develop a customized protocol for your target protein after estimating endotoxin removal efficiency and sample recovery. We will use a combination of the following methods to ensure maximum efficiency.
The hydrophobic column interacts with nonpolar protein surfaces through van der Waals forces for endotoxin removal.
It uses composite polyacrylamide as the column, which is highly porous. Ultrafiltration results in relatively good endotoxin clearance from product solutions when the products are of low molecular weight.
The high selectivity of affinity chromatography eliminates the need for multiple purification steps and reduces production costs. At the same time, this method can remove endotoxin with high specificity and efficiency, resulting in excellent target recovery.
Creative BioMart offers a corresponding endotoxin removal service. You can purchase the corresponding endotoxin removal kit and related accessory products according to the needs of your own samples. We guarantee that all instruments, water, reagents, and consumables used in the experiment are free of endotoxins, and the experiment is conducted in a clean room to ensure that low levels of endotoxins are returned to the sample. In addition, we can also provide you with related services including:
|Project name||Endotoxin removal services in plasmid DNA|
|Removal cycle||5-7 days.|
|Removal purpose||Endotoxin content is usually very high, and environmental or operational factors can easily lead to LPS contamination of products. LPS has a very strong heat source. Very small amounts (ng content) can cause high fever, diarrhea, vasodilation, and even fainting or death. Therefore, the removal of LPS has always been an important step in biotechnology drug research.|
|Service including||Creative BioMart can screen for the source of endotoxin contamination in the production process and propose several solutions to remove endotoxin to reduce the endotoxin content in the finished product.|