A kind of protein solution bacterial endotoxin removal method

Abstract

The invention discloses a protein solution bacterial endotoxin removal method, which comprises the following steps: S1. Aluminum hydroxide gel preparation: adding 2mol / L sodium hydroxide to 4.5% aluminum chloride solution, the pH is 7.0-7.5, React at 2°C to 8°C for 1 to 2 hours, and collect the gel by centrifugation at 4000rpm; S2. Remove bacterial endotoxin in the protein solution: add the aluminum hydroxide gel prepared in step S1 to the protein solution, Stir at 100 rpm for 1.5 to 2.5 hours at °C, and centrifuge at 4000 rpm to collect the supernatant after removing endotoxin. In the present invention, the adsorption of protein by aluminum hydroxide gel is mainly electrostatic adsorption, which can reduce or prevent the adsorption of protein by aluminum hydroxide in the presence of phosphate and high-concentration NaCl (both contained in PB buffer solution). Can remove 99% of bacterial endotoxins.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for removing bacterial endotoxin from a protein solution. Background technique [0002] Bacterial endotoxin (Endotoxin) is a unique structure on the cell wall of Gram-negative bacteria. Endotoxin is an exogenous pyrogen. The main chemical component of bacterial endotoxin is lipopolysaccharide (LPS), and its toxic components are mainly lipids. A. Endotoxin is heat-resistant and stable, and its activity can only be destroyed at 180°C for 2 hours. The toxic effects of endotoxins of various bacteria are weak and roughly the same, which can cause fever, endotoxin shock and disseminated intravascular coagulation. Usually, LPS exists in the complete form on the outer membrane of bacterial cells. When it is released from the bacteria, its activity is 50-100 times that of endotoxin on the cell wall. It has a wide range of biological activities and can cause severe inflam...

AI Technical Summary

Problems solved by technology

Even after the purification process, the bacterial endotoxin has been reduced, but the bacterial endotoxin content is still very high, which is the main reason for the vaccine's febrile reaction

Most antibiotics used clinically in the treatment of bacterial infections (mainly Gram-negative bacterial infections) lead to the release of a large number of bacterial endotoxins due to bactericidal / bacteriostatic reasons, making the condition worse

[0004] At present, the removal methods of bacterial endotoxin in the solution include: (1) Ultrafiltration technology separates the target substance and bacterial endotoxin by the difference in molecular size. Effective, suitable for small molecular substances; (2) acid-base chemical treatment is suitable for object treatment, usually not suitable for protein solutions; (3) chromatography (ie ion exchange, hydrophobic) equipment is expensive, material consumption is large, the operation is complicated, and Special chromatographic media need to be customized; (4) Positive charge filter filtration is usually used for small molecular substances and water treatment, not suitable for protein substances