What is endotoxin spiking?
Endotoxins are cell wall components of Gram-negative bacteria. A release of endotoxins into the human blood stream results in an inflammation reaction that can lead to life-threatening conditions like sepsis. Within these studies, endotoxin is added (spiked) to an undiluted product.
What is endotoxin removal?
Endotoxin is continuously shed from the outer membrane of viable gram-negative bacteria and is released when the bacterial cell dies. Although bacteria are often removed by using a 0.2 μm sterilizing grade filter, LPS itself is difficult to remove or inactivate because it is extremely heat and pH stable.
What is the purpose of endotoxins?
Endotoxins are found in the outer membrane of the cell wall of Gram-negative bacteria. They elicit a strong immune response in man (e.g., fever, septic shock), and cannot be removed from materials by normal sterilization processes.
What is the purpose of endotoxin testing?
What is Endotoxin Testing? LAL (Limulus amebocyte lysate) endotoxin testing is an in-vitro assay used for the detection and quantitation of bacterial endotoxins in injectable products or implantable medical devices that will make direct or indirect contact with the bloodstream or spinal fluid.
What is endotoxin spike recovery?
Spike recovery: The spike is an aliquot of test sample together with a known amount of endotoxin. The spike serves as the check for interference (inhibition and enhancement). Inhibition and enhancement are conditions that adversely alter the recovery of endotoxin in a test sample.
How do you filter out endotoxins?
Traditionally, endotoxins have been removed by reverse osmosis or ultrafiltration membranes of a 10,000 NMWL cutoff12. However, this approach is not practical when a component of a drug product has a molecular weight greater than 10 kDa.
Why is it called endotoxin?
LPS is also called an endotoxin because it is a toxin located inside the bacterial cell. It was originally theorized that endotoxin is released once the bacteria dies. LPS is structurally divided into three main parts, the O-antigen, the Core oligosaccharide and Lipid-A.
What is endotoxin test?
The bacterial endotoxins test (BET) is a test to detect or quantify endotoxins from Gram- negative bacteria using amoebocyte lysate from the horseshoe crab (Limulus polyphemus or. Tachypleus tridentatus).
How do you test for endotoxin?
Test procedure: A BET involves analyzing the liquid sample or sample extract using Limulus Amebocyte Lysate (LAL). LAL is a reagent made from the blood of the horseshoe crab. In the presence of bacterial endotoxins, the lysate reacts to form a clot or cause a color change depending on the technique.
What are endospores used for?
An endospore is a dormant, tough, non-reproductive structure produced by a small number of bacteria from the Firmicute family. The primary function of most endospores is to ensure the survival of a bacterium through periods of environmental stress.
How do Lipopolysaccharides work?
LPS is the most abundant antigen on the cell surface of most Gram-negative bacteria, contributing up to 80% of the outer membrane of E. coli and Salmonella. LPS increases the negative charge of the cell membrane and helps stabilize the overall membrane structure.