Recombinant protein is a protein obtained by using genetic recombination technology. First, a recombinant vector is introduced, a gene fragment that can be translated into a target protein is ligated to the recombinant vector, and then transferred to a host cell that can express the target protein to express a specific recombinant protein molecule. Currently, the production of recombinant proteins mainly includes four major systems: prokaryotic protein expression, mammalian cell protein expression, yeast protein expression, and insect cell protein expression. As a biopharmaceutical product, the resulting recombinant protein plays an important role in medicine. After purification of the recombinant vector plasmid, endotoxin may adversely affect some downstream applications. If the endotoxin level is high enough, it will affect the transfection process of the cell's DNA, resulting in reduced transfection efficiency.
Endotoxin is widely found in the cell walls of Gram-negative bacteria, such as E. coli. In the process of expressing recombinant protein, E. coli produces macroendotoxin. Endotoxin, as a heat source, leads to inflammatory and febrile reactions in the organism, so when the recombinant protein is used in some animal experiments, endotoxin needs to be removed in order to avoid causing adverse reactions in the animals and thus affecting the experimental results. Endotoxin may also be used for gene therapy of polypeptides and antibodies produced by recombinant proteins Applications have an adverse effect because they can cause side effects such as fever in animals and humans and endotoxemia, leading to misunderstandings of research results. In summary, it is of great significance to detect and remove endotoxin in recombinant proteins.
Endotoxin control and removal is based on the principle of endotoxin-binding peptide purification resin, which needs to be controlled from the beginning of protein preparation experiments. Practical utensils, instruments, water, reagents, and pipettes are used for endotoxin removal. At the same time, the experiments were performed in a clean room, ensuring that all levels of low endotoxin tests were achieved.
Fig1. The general process of endotoxin removal.
Creative BioMart offers a variety of endotoxin removal services covering all existing endotoxin removal methods to meet the specific needs of different customers.
Removal of endotoxin with an affinity adsorbent that selectively adsorbs endotoxin has the significant advantage of high protein recovery (nearly 100%). Endotoxin from different strains has different chemical structures, which requires different structures and functions of affinity ligands. In addition, since the endotoxin concentration in the protein solution is low, a strong interaction force between the endotoxin and the affinity ligand is required.
Endotoxin in solution usually aggregates to form aggregates with a molecular weight greater than 300KDa. When the molecular weight of the recombinant protein is small, an ultrafiltration membrane with a molecular weight greater than 100 KDa can be used to remove endotoxin from the protein solution.
Table.1 Comparison of our endotoxin removal methods
|Affinity chromatography||Specific adsorption||
|Ultrafiltration||A small molecular weight of the target product||
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 recombinant protein|
|Testing cycle||5-7 days.|
|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.|