The venom of Crotalus durissus terrificus (Cdt) is a source of a wide variety of toxins, some of them with interesting pharmacological applications. Of these toxins, the phospholipase A2 (PLA2) subunit of crotoxin (Ctx) has been studied for its potential as an antiviral and antibacterial agent. Peptides have proven useful ligands for the purification of numerous molecules, including antibodies, toxins, enzymes and other proteins. Here, we sought to use a phosphopeptide (P-Lys) as a ligand for PLA2 purification. P-Lys was synthesized in solid phase on Rink-Amide-ChemMatrix resin, immobilized on NHS-agarose, and then evaluated as a chromatographic matrix. Under the best conditions, total protein adsorption reached 39% and only the eluate fraction presented PLA2 activity. Analysis of the eluate by SDS-PAGE showed three bands, one corresponding to the molecular weight of PLA2 (14 kDa). Said bands were analyzed by mass spectrometry and identified as PLA2 and its multimers. The final product showed a purity of over 90%. In addition, slightly changing the process conditions also allowed the isolation of crotamine.

中文翻译：

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通过亲和色谱法使用磷酸肽作为配体从可怕的Cratarus durisuss terrificus毒液中纯化磷脂酶A2。

猪屎豆（Ctalt durissus terrificus，Cdt）的毒液是多种毒素的来源，其中一些具有有趣的药理应用。在这些毒素中，已经研究了Crotoxin（Ctx）的磷脂酶A2（PLA2）亚基作为抗病毒和抗菌剂的潜力。肽已被证明可用于多种分子的纯化，包括抗体，毒素，酶和其他蛋白质。在这里，我们试图使用磷酸肽（P-Lys）作为PLA2纯化的配体。在Rink-Amide-ChemMatrix树脂上固相合成P-Lys，固定在NHS-琼脂糖上，然后评估为色谱基质。在最佳条件下，总蛋白吸附达到39％，只有洗脱液部分具有PLA2活性。通过SDS-PAGE分析洗脱液显示出三个谱带，一个对应于PLA2的分子量（14 kDa）。所述条带通过质谱分析并且鉴定为PLA 2及其多聚体。最终产物显示出超过90％的纯度。另外，稍微改变工艺条件也可以分离出巴豆胺。