Division of BioAnalytical Chemistry, Amsterdam Institute of Molecular and Life Sciences, Vrije Universiteit Amsterdam, De Boelelaan 1085, 1081 HV Amsterdam, The Netherlands.
Centre for Analytical Sciences Amsterdam (CASA), 1098 XH Amsterdam, The Netherlands.
Toxins (Basel). 2020 Jan 16;12(1):53. doi: 10.3390/toxins12010053.
Venomous snakebite is one of the world's most lethal neglected tropical diseases. Animal-derived antivenoms are the only standardized specific therapies currently available for treating snakebite envenoming, but due to venom variation, often this treatment is not effective in counteracting all clinical symptoms caused by the multitude of injected toxins. In this study, the coagulopathic toxicities of venoms from the medically relevant snake species , , , , and were assessed. The venoms were separated by liquid chromatography (LC) followed by nanofractionation and parallel mass spectrometry (MS). A recently developed high-throughput coagulation assay was employed to assess both the pro- and anticoagulant activity of separated venom toxins. The neutralization capacity of antivenoms on separated venom components was assessed and the coagulopathic venom peptides and enzymes that were either neutralized or remained active in the presence of antivenom were identified by correlating bioassay results with the MS data and with off-line generated proteomics data. The results showed that most snake venoms analyzed contained both procoagulants and anticoagulants. Most anticoagulants were identified as phospholipases As (PLAs) and most procoagulants correlated with snake venom metalloproteinases (SVMPs) and serine proteases (SVSPs). This information can be used to better understand antivenom neutralization and can aid in the development of next-generation antivenom treatments.
蛇伤是世界上最致命的被忽视热带病之一。动物来源的抗蛇毒血清是目前可用于治疗蛇咬伤中毒的唯一标准化特效疗法，但由于毒液的变异，这种治疗方法常常无法有效对抗多种注入毒素引起的所有临床症状。在这项研究中，评估了来自医学相关蛇种 、 、 、 、 和 的毒液的凝血毒性。毒液通过液相色谱（LC）分离，然后进行纳滤和并行质谱（MS）分析。采用最近开发的高通量凝血测定法评估分离的毒液毒素的促凝和抗凝活性。评估抗蛇毒血清对分离的毒液成分的中和能力，并通过将生物测定结果与 MS 数据和离线生成的蛋白质组学数据相关联，鉴定出中和或在抗蛇毒血清存在下仍保持活性的凝血性毒液肽和酶。结果表明，大多数分析的蛇毒液既含有促凝剂也含有抗凝剂。大多数抗凝剂被鉴定为磷脂酶 A（PLA），而大多数促凝剂与蛇毒金属蛋白酶（SVMP）和丝氨酸蛋白酶（SVSP）相关。这些信息可用于更好地了解抗蛇毒血清的中和作用，并有助于开发下一代抗蛇毒血清治疗方法。
