Use of an Antimicrobial Protein for Endotoxin Detection in a Competitive Electrochemical Assay

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• 作者：Graciela Priano and Fernando Battaglini
• 刊名：Analytical Chemistry
• 出版年：2005
• 出版时间：August 1, 2005
• 年：2005
• 卷：77
• 期：15
• 页码：4976 - 4984
• 全文大小：293K
• 年卷期：v.77,no.15(August 1, 2005)
• ISSN：1520-6882

文摘

Endotoxins, also referred to as pyrogens, are part of thecellular walls of Gram-negative bacteria and are capableof inducing fever when entering into the blood stream.Current methods of detection involve the use of eitheramoebocytes from horseshoe crab that clots upon exposure to endotoxin or a live assay on rabbits. In this work,the detection of endotoxins by an electrochemical competitive assay is presented. Two configurations of modifiedelectrodes were constructed using a recombinant endotoxin neutralizing protein (ENP) as recognition element.A modified lipopolysaccharide with horseradish peroxidase was used for the competitive assay. Modified electrodes constructed by electrostatic interaction of ENP andan electroactive polymer can detect the presence ofendotoxins in concentrations as low as 0.2EU mL^-1,below the limit imposed for water in injectable drugs inthe American Pharmacopoeia. Modified electrodes constructed by covalent linking of ENP to a carboxymethyldextran matrix bound to the electrode show a betterdynamic range but a higher detection limit.