Current technologies to endotoxin detection and removal for biopharmaceutical purification

Abstract

Endotoxins are the major contributors to the pyrogenic response caused by contaminated pharmaceutical products, formulation ingredients, and medical devices. Recombinant biopharmaceutical products are manufactured using living organisms, including Gram‐negative bacteria. Upon the death of a Gram‐negative bacterium, endotoxins (also known as lipopolysaccharides) in the outer cell membrane are released into the lysate where they can interact with and form bonds with biomolecules, including target therapeutic compounds. Endotoxin contamination of biologic products may also occur through water, raw materials such as excipients, media, additives, sera, equipment, containers closure systems, and expression systems used in manufacturing. The manufacturing process is, therefore, in critical need of methods to reduce and remove endotoxins by monitoring raw materials and in‐process intermediates at critical steps, in addition to final drug product release testing. This review paper highlights a discussion on three major topics about endotoxin detection techniques, upstream processes for the production of therapeutic molecules, and downstream processes to eliminate endotoxins during product purification. Finally, we have evaluated the effectiveness of endotoxin removal processes from a perspective of high purity and low cost.

Central Nervous System Delivery of Antibodies and Their Single-Domain Antibodies and Variable Fragment Derivatives with Focus on Intranasal Nose to Brain Administration

“…One approach to reduce the prices of biopharmaceuticals is to produce smaller and less complex molecules by using cheaper production systems, e.g., yeast or bacteria instead of mammalian cells [ 24 , 25 ]. However, bacteria are not optimal for industrial production scales, as bacteria-derived endotoxins carry a high risk of immune reactions [ 26 ].…”

Human Platelet Lysate for Good Manufacturing Practice-Compliant Cell Production

“…Endotoxins are toxic breakdown products of Gram-negative bacteria. When inoculated into the blood stream, endotoxins can trigger a severe pyrogenic reaction, activation of the coagulation cascade, hypotension and shock [ 85 ]. A maximum threshold of endotoxin concentration has to be defined depending on product type and route of administration.…”

“…In the chromogenic method, the natural substrate is replaced by a chromogen-coupled substrate. The chromogenic molecule is released after substrate cleavage by an endotoxin-activated enzyme [ 81 , 85 ]. Recently, the use of recombinant factor C was published as an alternative to the classic LAL method [ 86 ].…”

ClearColi as a platform for untagged pneumococcal surface protein A production: cultivation strategy, bioreactor culture, and purification

“…LPS may cause an endotoxin response in humans, even at extremely low concentrations (Taguchi et al 2015 ). The LPS structure consists of three regions: a core polysaccharide, an external long-chain polysaccharide, and a nonpolar lipid (lipid A) that is the portion of the molecule responsible for toxicity (Schneier et al 2020 ). Lipid A binds toll-like receptor 4 (TLR4), triggering the production of proinflammatory cytokines and activating the coagulation cascade, which can lead to septic shock and death (Ramachandran 2014 ).…”