Tevell Åberg Annica, Björnstad Kristian, Hedeland Mikael
Annica Tevell Åberg, PhD, is a Senior Researcher; Kristian Björnstad, PhD, is a Senior Researcher; and Mikael Hedeland, PhD, is an Associate Professor and Deputy Head of Department; all at the Department of Chemistry, Environment and Feed Hygiene, National Veterinary Institute (SVA) , Uppsala, Sweden. Dr. Åberg and Dr. Hedeland are also affiliated with the Division of Analytical Pharmaceutical Chemistry, Uppsala University, Uppsala, Sweden .
Biosecur Bioterror. 2013 Sep;11 Suppl 1:S215-26. doi: 10.1089/bsp.2012.0072.
This review focuses on mass spectrometric detection of protein-based toxins, which are among the most toxic substances known. Special emphasis is given to the bacterial toxins botulinum neurotoxin from Clostridium botulinum and anthrax toxins from Bacillus anthracis as well as the plant toxin ricin produced by Ricinus communis. A common feature, apart from their extreme toxicity, is that they are composed of 2 polypeptide chains, one of which is responsible for cell uptake and another that has enzymatic function with the ability to destroy basic cellular functions. These toxins pose a threat, both regarding natural spread and from a terrorism perspective. In order for public health and emergency response officials to take appropriate action in case of an outbreak, whether natural or intentional, there is a need for fast and reliable detection methods. Traditionally, large molecules like proteins have been detected using immunological techniques. Although sensitive, these methods suffer from some drawbacks, such as the risk of false-positives due to cross-reactions and detection of inactive toxin. This article describes recently developed instrumental methods based on mass spectrometry for the reliable detection of botulinum neurotoxins, anthrax toxins, and ricin. Unequivocal identification of a protein toxin can be carried out by mass spectrometry-based amino acid sequencing. Furthermore, in combination with antibody affinity preconcentration and biochemical tests with mass spectrometric detection demonstrating the toxin's enzymatic activity, very powerful analytical methods have been described. In conclusion, the advent of sensitive, easily operated mass spectrometers provides new possibilities for the detection of protein-based toxins.
本综述聚焦于基于蛋白质的毒素的质谱检测，这些毒素是已知毒性最强的物质之一。特别强调了来自肉毒杆菌的肉毒杆菌神经毒素、来自炭疽杆菌的炭疽毒素以及蓖麻毒素，后者是由蓖麻产生的植物毒素。除了具有极高的毒性外，这些毒素的一个共同特征是它们由两条多肽链组成，其中一条负责细胞摄取，另一条具有酶功能，能够破坏基本的细胞功能。无论是从自然传播还是从恐怖主义角度来看，这些毒素都构成了威胁。为了让公共卫生和应急响应官员在爆发事件（无论是自然爆发还是人为蓄意）发生时能够采取适当行动，需要快速且可靠的检测方法。传统上，像蛋白质这样的大分子是使用免疫技术进行检测的。尽管这些方法很灵敏，但也存在一些缺点，比如由于交叉反应导致假阳性的风险以及对无活性毒素的检测。本文介绍了最近基于质谱开发的仪器方法，用于可靠检测肉毒杆菌神经毒素、炭疽毒素和蓖麻毒素。基于质谱的氨基酸测序能够明确鉴定蛋白质毒素。此外，结合抗体亲和预浓缩以及通过质谱检测证明毒素酶活性的生化测试，已经描述了非常强大的分析方法。总之，灵敏、易于操作的质谱仪的出现为基于蛋白质的毒素的检测提供了新的可能性。
