Lipopolysaccharide,LPSremoval, Depyrogenation

Abstract

Advances in biotechnology have opened up numerous possibilities for the large‐scale production of many biomolecules that are important for research, pharmaceuticals, and industrial applications. The development of techniques and methods for the separation and purification of proteins and other biomolecules has been of paramount importance for many of these advances in the biotechnology industry. Such biomolecules may be expressed as soluble extracellular form, soluble intracellular form, or as membrane constitute or inclusion bodies. In all three cases, use of cellular rupture procedures for the recovery of the biomolecules of interest, which lead to the release of a large quantity of lipopolysaccharide (LPS)‐type endotoxins, is necessary. The presence of small amounts of endotoxin in recombinant protein preparations can cause side effects such as endotoxin shock, tissue injury, and even death in host organism. Owing to these reactions, research on the methods used to remove endotoxins from drugs, injectables, and other biological and pharmaceutical products is necessary. Taking into consideration the main techniques used as well as the state of the art of the technology in this field, the techniques of endotoxin determination and interaction of endotoxins with proteins are presented.

A trimeric coiled-coil motif binds bacterial lipopolysaccharides with picomolar affinity

“…It is therefore imperative that especially pharmaceutical products are completely LPS-free. Due to the omnipresence of bacteria, and hence, LPS, in the environment, this is difficult to achieve, and requires both highly sensitive tests and challenging LPS removal efforts ( Magalhães and Pessoa, 2010 ). Current methods for LPS removal include for example affinity chromatography with Polymyxin B-based resins that specifically bind LPS; the gold standard for LPS detection today still is the Limulus amoebocyte lysate (LAL) assay that relies on the blood of an endangered species, the horseshoe crab ( Schneier et al., 2020 ).…”