Synthesis and Application of a Novel Ligand for Affinity Chromatography Based Removal of Endotoxin from Antibodies

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• 作者：Laura A. McAllister ; Mark S. Hixon ; Roberta Schwartz ; Diane S. Kubitz ; Kim D. Janda
• 刊名：Bioconjugate Chemistry
• 出版年：2007
• 出版时间：March 2007
• 年：2007
• 卷：18
• 期：2
• 页码：559 - 566
• 全文大小：195K
• 年卷期：v.18,no.2(March 2007)
• ISSN：1520-4812

文摘

Endotoxin or lipopolysaccharide (LPS) contamination in proteins expressed by Gram-negative bacteria is a majordrawback associated with protein expression. Endotoxin intoxication in humans and animals above a certainthreshold level can result in a fatal immune response. Reduction in endotoxin levels is therefore essential beforeproteins can be used in in vivo studies or sold as pharmaceutical products. Affinity chromatography employingthe peptide Polymyxin B (PMB) as an affinity ligand is one way in which endotoxin contamination has beenaddressed; this is, however, a costly process. We describe the synthesis of a novel affinity ligand based on thestructure of the drug pentamidine, which can be applied effectively in endotoxin removal. The synthetic route tothis ligand is straightforward and inexpensive, while the ligand can be readily immobilized onto activated sepharosebeads. Thus, we demonstrate that these pentamidine affinity beads bind endotoxin/LPS with comparable capacityto PMB affinity systems, that the beads can be recycled efficiently and economically without loss of bindingcapacity, and application of the functionalized beads for endotoxin removal in an authentic contaminated antibodysample.