Heterologous expression of four recombinant toxins from Panamanian scorpions of the genus Tityus and Centruroides for production of antivenom

IF 3.6 Q2 TOXICOLOGY
Marcos H. Salazar , Herlinda Clement , Ligia L. Corrales-García , Jairo Sánchez , John Cleghorn , Fernando Zamudio , Lourival D. Possani , Hildaura Acosta , Gerardo Corzo
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引用次数: 3

Abstract

Background

The development of more effective antivenoms remains a necessity for countries where scorpionism is a public health problem. Also, the regionalization of antivenoms may be important for some countries with special scorpionism characteristics.

Objective

Production of antibodies capable of neutralizing the lethal effect of the venom of three scorpion species from Panama.

Methods

The primary structures of two neurotoxins from T. pachyurus, one from T. cerroazul and another from C. bicolor were elucidated using N-terminal amino acid degradation and Sanger gene cloned sequencing. The obtained mRNA transcripts were cloned and expressed using E. coli vectors. Different bacterial expression conditions were tested and the best culture conditions for each expressed protein is reported. The expressed scorpion toxins were purified by chromatographic methods and used as immunogens in rabbits.

Results

The antibodies produced under the reported immunization scheme show better neutralization (ED50) than other reported commercial antivenoms used to neutralize similar species scorpion venoms under similar LD50 conditions.

Conclusion

The information reported here shows the proof of concept for selecting recombinant immunogens with the ability to produce antibodies for neutralizing the lethal effects of the most important medical species of scorpions in Panama.

Abstract Image

四种巴拿马蝎属重组毒素的异源表达及抗蛇毒血清的制备
发展更有效的抗蛇毒血清对于那些蝎子病是公共卫生问题的国家来说仍然是必要的。此外,抗蛇毒血清的区域化可能对一些具有特殊蝎子病特征的国家很重要。目的制备能中和巴拿马三种蝎子毒液致死作用的抗体。方法采用n端氨基酸降解和Sanger基因克隆测序的方法,对两种不同神经毒素的主要结构进行分析。将获得的mRNA转录本克隆并利用大肠杆菌载体进行表达。试验了不同的细菌表达条件,并报道了每种表达蛋白的最佳培养条件。表达的蝎毒素经层析纯化后作为免疫原用于家兔。结果在相同的LD50条件下,该免疫方案产生的抗体比其他已报道的商业抗蛇毒血清具有更好的中和效果(ED50)。结论选择重组免疫原能够产生抗体来中和巴拿马最重要的药用蝎子的致死效应,这一概念得到了证明。
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来源期刊
Toxicon: X
Toxicon: X Pharmacology, Toxicology and Pharmaceutics-Toxicology
CiteScore
6.50
自引率
0.00%
发文量
33
审稿时长
14 weeks
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