Pyrogen Removal

Endotoxins are lipopolysaccharides (LPS) located on the cell wall (cell membranes) of gram negative bacteria. The presence of even small amounts of endotoxin in recombinant protein preparations when injected into patients may cause systemic inflammatory reactions running the spectrum from tissue injury, to endotoxin shock and death.

Pharmaceutical products produced using bacteria as cellular factories are virtually always contaminated with LPS and all measurable endotoxin must be removed during the production process. The maximum acceptable level of endotoxin for intravenous applications is set at 5 endotoxin units (EU) per kg of body weight per hour.

Specially activated alumina with surface modified chemical moieties has proven to provide a superior tool for the purification of endotoxins due to its amphoteric property. No other commercially available agent can provide such a rewarding pH response or offer a better or cost effective method for the removal of chemical pyrogens from a protein or peptide solution.

It is clear that endotoxins develop especially strong binding to adsorbents carrying positively charged functional groups. Electrostatic interactions play an important role during endotoxin adsorption. Proteins are also amphoteric. Since proteins are amphoteric molecules, electrostatic interactions are not as strong as for the mainly negatively charged endotoxin. Owing to the globular structure of proteins, charged and hydrophobic groups are fixed and cannot be twisted towards functional groups or surface structures of the adsorbents. Additional benefits provided through the use of activated alumina include low cost, limited safety issues, extremely well defined chemical characteristics and minimal impact on the bioactivity of protein when placed into a standard manufacturing process.

The removal of pyrogens using specially designed activated alumina can be performed using either column chromatography or batch treatment. When using column chromatography the final product is achieved by filling a column with the alumina modified to enhance pyrogen adsorbance, prewashing the packed column with a suitable buffer and then passing the pyrogen containing solution through the column. In the method of batch treatment the final product freed of pathogens can be obtained by stirring the pyrogen adsorbent in a pyrogen containing solution of the desired compound and then removing the adsorbent.

DynaPharma™ Pyrogen using either column chromatography or batch processing procedures has proven to be a superior tool for the isolation and removal of chemical pyrogens. The surface of activated alumina may be modified to help enhance pyrogen removal.

DynaPharma™ Pyrogen Product Info | Chemical Pyrogen Removal Through the use of Activated Alumina