Departments of Biology, Bioengineering, and Medicinal Chemistry, University of Utah, Salt Lake City, UT, USA.
SLAS Technol. 2020 Aug;25(4):397-403. doi: 10.1177/2472630320932890. Epub 2020 Jun 26.
Heparin has been in clinical use as an anticoagulant for the last eight decades and used worldwide in more than 100 million medical procedures every year. This lifesaving drug is predominantly obtained from ~700 million pig intestines or bovine organs through millions of small and medium-sized slaughterhouses. However, the preparations from animal sources have raised many safety concerns, including the contamination of heparin with potential pathogens, proteins, and other impurities. In fact, contaminated heparin preparations caused 149 deaths in several countries, including the United States, Germany, and Japan in 2008, highlighting the need for implementing sensitive and simple analytical techniques to monitor and safeguard the heparin supply chain. The contaminant responsible for the adverse effects in 2008 was identified as oversulfated chondroitin sulfate (OSCS). We have developed a very sensitive, facile method of detecting OSCS in heparin lots using a nanosensor, a gold nanoparticle-heparin dye conjugate. The sensor is an excellent substrate for heparitinase enzyme, which cleaves the heparin polymer into smaller disaccharide fragments, and therefore facilitates recovery of fluorescence from the dye upon heparitinase treatment. However, the presence of OSCS results in diminished fluorescence recovery from the nanosensor upon heparitinase treatment, because OSCS inhibits the enzyme. The newly designed nanosensor can detect as low as 1 × 10% (w/w) OSCS, making it the most sensitive tool available to date for the detection of trace amounts of OSCS in pharmaceutical heparins. In this report, we describe a simple methodology for the preparation of nanosensor and its application in the detection of OSCS contaminants.
肝素作为抗凝剂在临床上已经使用了近 80 年，每年在全球有超过 1 亿例医疗程序中使用。这种救命药物主要从 ~7 亿个猪肠或牛器官中通过数百万个中小屠宰场获得。然而，来自动物源的制剂引起了许多安全问题，包括肝素可能被潜在病原体、蛋白质和其他杂质污染。事实上，受污染的肝素制剂在包括美国、德国和日本在内的几个国家导致了 149 人死亡，这突出表明需要实施敏感和简单的分析技术来监测和保障肝素供应链的安全。2008 年导致不良反应的污染物被确认为过度硫酸化的软骨素硫酸盐（OSCS）。我们已经开发了一种非常灵敏、简便的方法，使用纳米传感器（一种金纳米粒子-肝素染料缀合物）检测肝素批次中的 OSCS。该传感器是肝素酶的极佳底物，肝素酶将肝素聚合物切割成较小的二糖片段，从而便于在肝素酶处理后从染料中恢复荧光。然而，OSCS 的存在会导致肝素酶处理后纳米传感器的荧光恢复减少，因为 OSCS 会抑制该酶。新设计的纳米传感器可以检测低至 1×10%（w/w）的 OSCS，使其成为迄今为止用于检测药物肝素中痕量 OSCS 的最灵敏工具。在本报告中，我们描述了一种制备纳米传感器的简单方法及其在 OSCS 污染物检测中的应用。
