Giraldo Luz Elena Romero, Pulido Sergio, Berrío Mario Andrés, Flórez María Fernanda, Rey-Suárez Paola, Núñez-Rangel Vitelbina, Córdoba Mónica Saldarriaga, Pereañez Jaime Andrés
Research Group in Toxinology, Pharmaceutical, and Food Alternatives, University of Antioquia, Medellín, Colombia.
LifeFactors Free Zone SAS, Rionegro, Antioquia, Colombia.
J Venom Anim Toxins Incl Trop Dis. 2024 Nov 25;30:e20230074. doi: 10.1590/1678-9199-JVATITD-2023-0074. eCollection 2024.
is a coral snake of public health concern in Colombia. Its venom is mainly composed of three-finger toxins (3FTxs), Mipartoxin-1 being the most abundant protein partially responsible for its lethal effect. In this work, we present the production of Mipartoxin-1 in a recombinant form and evaluate its immunogenic potential. A genetic construct HisrMipartoxin-1 was cloned into the pET28a vector and heterologous expression was obtained in BL21 (DE3). The recombinant HisrMipartoxin-1 protein was extracted from inclusion bodies, refolded , and isolated by affinity and RP-HPLC chromatography. The lethal effect of HisrMipartoxin-1 was tested, and antibodies against HisrMipartoxin-1 were produced by immunization in rabbits. The antibody titers were monitored by an ELISA test. The neutralizing ability of the antibodies, against the lethal effect of native toxins and venom, was also assessed. HisrMipartoxin-1 was detected on SDS-PAGE, with a molecular mass of around 11 kDa. The retention time was 16.0 minutes. HisrMipartoxin-1 did not exhibit lethality in mice; however, antibodies against HisrMipartoxin-1 recognized the native toxin, the whole venom of , and a 3FTx from another species within the genus. Furthermore, antibodies against HisrMipartoxin-1 completely neutralized the lethal effect of native Mipartoxin-1 in mice but not whole venom. These findings indicate that HisrMipartoxin-1 might be used as an immunogen to develop anticoral antivenoms or complement them. This work is the first report of the heterologous expression of 3FTx from .
是哥伦比亚一种受公共卫生关注的珊瑚蛇。其毒液主要由三指毒素(3FTxs)组成,米帕毒素 -1是最丰富的蛋白质,部分负责其致死作用。在这项工作中,我们展示了重组形式的米帕毒素 -1的生产,并评估了其免疫原性潜力。将基因构建体Hisr米帕毒素 -1克隆到pET28a载体中,并在BL21(DE3)中获得异源表达。从包涵体中提取重组Hisr米帕毒素 -1蛋白,进行重折叠,并通过亲和色谱和反相高效液相色谱进行分离。测试了Hisr米帕毒素 -1的致死作用,并通过在兔体内免疫产生了针对Hisr米帕毒素 -1的抗体。通过ELISA试验监测抗体滴度。还评估了抗体对天然毒素和毒液致死作用的中和能力。在SDS - PAGE上检测到Hisr米帕毒素 -1,分子量约为11 kDa。保留时间为16.0分钟。Hisr米帕毒素 -1在小鼠中未表现出致死性;然而,针对Hisr米帕毒素 -1的抗体识别天然毒素、该蛇的全毒液以及来自该属另一物种的一种3FTx。此外,针对Hisr米帕毒素 -1的抗体完全中和了小鼠体内天然米帕毒素 -1的致死作用,但未中和全毒液。这些发现表明Hisr米帕毒素 -1可作为免疫原用于开发抗珊瑚蛇毒血清或对其进行补充。这项工作是关于该蛇3FTx异源表达的首次报道。