Immunogenic potential and neutralizing ability of a heterologous version of the most abundant three-finger toxin from the coral snake Micrurus mipartitus.
Luz Elena Romero Giraldo, Sergio Pulido, Mario Andrés Berrío, María Fernanda Flórez, Paola Rey-Suárez, Vitelbina Núñez-Rangel, Mónica Saldarriaga Córdoba, Jaime Andrés Pereañez
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引用次数: 0
Abstract
Background:Micrurus mipartitus 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. Methods: A genetic construct HisrMipartoxin-1 was cloned into the pET28a vector and heterologous expression was obtained in E. coli BL21 (DE3). The recombinant HisrMipartoxin-1 protein was extracted from inclusion bodies, refolded in vitro, 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 M. mipartitus venom, was also assessed. Results: 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 M. mipartitus, and a 3FTx from another species within the Micrurus genus. Furthermore, antibodies against HisrMipartoxin-1 completely neutralized the lethal effect of native Mipartoxin-1 in mice but not M. mipartitus whole venom. Conclusion: 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 M. mipartitus.
期刊介绍:
Journal of Venomous Animals and Toxins including Tropical Diseases (JVATiTD) is a non-commercial academic open access publication dedicated to research on all aspects of toxinology, venomous animals and tropical diseases. Its interdisciplinary content includes original scientific articles covering research on toxins derived from animals, plants and microorganisms. Topics of interest include, but are not limited to:systematics and morphology of venomous animals;physiology, biochemistry, pharmacology and immunology of toxins;epidemiology, clinical aspects and treatment of envenoming by different animals, plants and microorganisms;development and evaluation of antivenoms and toxin-derivative products;epidemiology, clinical aspects and treatment of tropical diseases (caused by virus, bacteria, algae, fungi and parasites) including the neglected tropical diseases (NTDs) defined by the World Health Organization.