Removal of endotoxin from recombinant protein preparations

  Endotoxins are toxic protoplasms that are released when a bacteria dies, disintegrates, and grows. Many of these endotoxins are lipopolysaccharides, which reside in the outer membrane of gram-negative bacteria. The gram-negative cell wall is composed of a thin, inner layer of peptidoglycan and an outer membrane of phospholipids, lipopolysaccharide, and lipoprotein. The lipopolysaccharides are made up of a polysaccharide chain and a lipid, which is responsible for the toxic effects. The toxins are continuously shed from the outer membrane of bacteria, and released into the environment. When producing recombinant proteins, these unwanted by products must be removed from the final product, especially when developing manufacturing processes.

The innate toxicity of endotoxins makes their removal an important step for the proteins’ application in several biological assays and for safe release of product. Since there are limits to the amounts of endotoxins allowed in human products, false readings in cell-based assays, caused by endotoxins, can cause serious issues, and so endotoxin removal techniques can be used to bind to the toxins and remove them from the culture. This can be especially important in the manufacturing of bio-pharmaceuticals, which are produced from living organisms such as bacteria. These toxins can become a serious problem in  bacterial infection, or when bacteria contaminate medications, food, and lab samples, because the toxins tend to resist heat and many other sterilization methods. As a result, when someone consumes a product contaminated with bacteria, which contain endotoxins, they can become sick. Just a small amount of endotoxin has been shown to produce fever, a decrease in blood pressure, and activation of inflammation in humans.

Endotoxin Removal is vital to producing a purified protein preperation. When it comes to the life science and pharmaceutical communites, Many companies outsource this process, along with many other stages of the molecular research phase.