A Heterologous Multiepitope DNA Prime/Recombinant Protein Boost Immunisation Strategy for the Development of an Antiserum against Micrurus corallinus (Coral Snake) Venom

Abstract

BackgroundEnvenoming by coral snakes (Elapidae: Micrurus), although not abundant, represent a serious health threat in the Americas, especially because antivenoms are scarce. The development of adequate amounts of antielapidic serum for the treatment of accidents caused by snakes like Micrurus corallinus is a challenging task due to characteristics such as low venom yield, fossorial habit, relatively small sizes and ophiophagous diet. These features make it difficult to capture and keep these snakes in captivity for venom collection. Furthermore, there are reports of antivenom scarcity in USA, leading to an increase in morbidity and mortality, with patients needing to be intubated and ventilated while the toxin wears off. The development of an alternative method for the production of an antielapidic serum, with no need for snake collection and maintenance in captivity, would be a plausible solution for the antielapidic serum shortage.Methods and FindingsIn this work we describe the mapping, by the SPOT-synthesis technique, of potential B-cell epitopes from five putative toxins from M. corallinus, which were used to design two multiepitope DNA strings for the genetic immunisation of female BALB/c mice. Results demonstrate that sera obtained from animals that were genetically immunised with these multiepitope constructs, followed by booster doses of recombinant proteins lead to a 60% survival in a lethal dose neutralisation assay.ConclusionHere we describe that the genetic immunisation with a synthetic multiepitope gene followed by booster doses with recombinant protein is a promising approach to develop an alternative antielapidic serum against M. corallinus venom without the need of collection and the very challenging maintenance of these snakes in captivity.

High-throughput immuno-profiling of mamba (Dendroaspis) venom toxin epitopes using high-density peptide microarrays

“…The potential of harnessing high-density peptide microarrays for identification of important toxin epitopes involved in toxin function and neutralisation by antivenoms may be utilised for design of next generation antivenoms based on DNA immunisation and immunisation with synthetic epitope strings13. DNA immunisation has been explored by a number of research groups424344454647, as has the use of synthetic epitope strings in the field of spider and scorpion antivenoms48.…”

“…Recently, studies involving immunoassay quantification of antivenom binding to immobilised synthetic peptides (SPOT synthesis), corresponding to individual segments of the amino acid sequence of a given toxin, have added valuable molecular insight by elucidating which sequences contain linear elements of epitopes recognised by given antivenoms67121314. These kind of meticulous epitope mapping experiments have also been performed on toxins from Tityus scorpions151617 and Loxosceles spiders18.…”

A synthetic biology approach for consistent production of plant‐made recombinant polyclonal antibodies against snake venom toxins

“…The use of immunization with DNA or with recombinantexpressed epitopes of relevant toxins is an example of this, aimed at circumventing the shortage of antigens in species with low venom yield, such as coral snakes (Ramos et al, 2016). In comparison, fully recombinant strategies can provide a more satisfactory solution in the long term, addressing not only the shortage of antigens, but also other problems arising from current therapies such as adverse reactions and reproducibility .…”

Peer Review #1 of "Exploration of immunoglobulin transcriptomes from mice immunized with three-finger toxins and phospholipases A2 from the Central American coral snake, Micrurus nigrocinctus (v0.1)"

“…Manuscript to be reviewed in overall toxicity have been identified, these being 3FTxs and PLA 2 s (Rey-Suárez et al, 2012;Vergara et al, 2014;Fernández et al, 2015;Castro et al, 2015;Ramos et al, 2016).…”