The pyrogen in the vaccine is mainly bacterial endotoxin, which is a lipopolysaccharide substance in the outer wall layer of Gram-negative bacteria. Such substances have heat resistance and chemical stability and cannot be easily destroyed. The traditional pyrogen removal method is an activated carbon adsorption method, but its applicability is limited because some active ingredients are also adsorbed. At present, the pyrogen content is measured by the rabbit method and the limulus reagent gel method. Various methods are used to remove bacterial endotoxins from vaccines.
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.
Each sample drug solution was appropriately diluted with bacterial endotoxin test water as a test solution (NPC). At the same time, this multiple dilution solution was made into a solution containing endotoxin standard 0.5EU/ml as a positive control (PPC). Add 0.1ml of the above solution to the reaction tube with 0.1ml limulus reagent added beforehand, mix them to detect the content of bacterial endotoxin, and repeat 2 tubes at each concentration.
Bacterial endotoxins are released during the growth and death of bacteria. After the cells rupture, a large amount of LPS is released into the solution. Endotoxin usually has a capsule-like structure, and its relative molecular weight, charge and hydrophobicity are similar to plasmid DNA, which makes it very difficult to purify vaccines. In addition, the presence of endotoxin seriously affects the transfection efficiency of cells. Therefore, bacterial endotoxin must be removed before vaccination, or serious consequences may result.
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 vaccine|
|Removal cycle||5-7 days.|
|Removal purpose||The pyrogen in the vaccine is mainly bacterial endotoxin, which is a lipopolysaccharide substance in the outer wall layer of Gram-negative bacteria. The endotoxin vaccine will cause many adverse reactions, so it needs to be removed before use.|
|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.|