Lipopolysaccharides Case Study

Q1
Associated Risks:
The endotoxins otherwise known as lipopolysaccharides are a serious source of concern in the biopharmaceutical industry. Endotoxins are not only a major source of product contamination, but they also pose a threat to the safety and wellbeing of the people involved in the manufacturing operations. One of the greatest risks associated the lipopolysaccharides is their ability to stimulate pyrogenic reactions in parenteral products. A very small of an endotoxin may prove extremely dangerous to the individual to whom the drug is being administered. As little as one endotoxin unit or 0.1 ng per a kilogram of the body weight, can lead to serious complications and the death of a patient.
General Characteristics:

Lipopolysaccharides

This as such may be considered a limitation. The use of ultrafiltration as a means of separation for highly viscous products may prove inefficient due to low filtration rates and therefore longer processing time. Another crucial factor with regards to ultra-filtration is the high cost of the system and associated modules. Another limitation is the size ratio of the proteins and the associated endotoxins. For ultra-filtration to be employed it is necessary that the contaminants are at least 25 larger than the molecules of interest. As a result of the size and the weight limitations of tangential flow filtration, it may only be used to filter out lipopolysaccharides from API’s and small

The first affinity tag was described in 1983 (Mathias Uhlén, 2008). It separates the proteins from endotoxins basing on the reversible interactions between the proteins and a matrix coupled ligand. It delivers excellent selectivity and resolution, provided that a ligand is available that can bind onto to the material of interest. It is believed that either the electrostatic or hydrophobic interactions are a result of the interactions between the specific ligand and the protein of interest. Desorption can be achieved by introducing a competitive ligand, by varying the pH or by changing the polarity of the matrix (GE Healthcare). While Affinity chromatography itself might not be exactly a novel technique, a number of surrounding technologies have been developed that make it such. A German company Hyglos, provides a resin known as EndoTrap HD. The resin is based on a bacteriophage-derived protein, which exhibits excellent binding characteristics. EndoTrap is very stable in comparison to other solutions and is non-toxic. The protein in question couples with the conserved region of the inner core of the endotoxin which makes it suitable for a wide range of applications (Hyglos,