The three-finger toxins (3FTxs) represent an extremely diverse protein family in elapid venoms, where the short chain α-neurotoxins are the most relevant toxin group from the clinical point of view. Essentially, the 3FTxs variability and the low proportions of α-neurotoxins in the venoms of North American coral snakes make it difficult to obtain effective elapid antivenoms against the envenomation symptoms caused mainly by these α-neurotoxins. In this work, thirty 3FTx transcript sequences were obtained from the venom glands of four coral snake species from Mexico (M. diastema, M. laticollaris, M. browni and M. tener). The transcripts were mined using a forward oligonucleotide based on the highly conserved signal peptide from the 3FTxs, and four of these transcripts, named MlatA1, B.D, B.E and D.H, encoded for short-chain α-neurotoxins. Additionally, one isoform of the D.H α-neurotoxin transcript was identified in the venom of M. diastema. The mature α-neurotoxin coded in the D.H transcript was heterologously expressed, and it was found soluble (4.2 mg/l) in the cytoplasm of a bacterial system. The recombinant D.H (rD.H) had an IC₅₀ value of 31.5 ± 4.4 nM on nicotinic acetylcholine receptors of the muscular type expressed in rhabdomyosarcoma cells (TE671). The rDH also had an LD₅₀ of 0.15 mg/kg mice, and it was evaluated as a potential immunogen in New Zealand rabbits. The protective capacity of rabbit sera was tested against two native coral snake α-neurotoxins, and against rD.H. One of the anti-rD.H rabbit sera was able to neutralize the lethality of all three neurotoxins when tested in groups of CD1 mice. This work contributes to the increasing understanding of the high diversity of 3FTxs, and shows that recombinant coral snake α-neurotoxins are promising supplements for hyperimmunization protocols for coral snake antivenom production.

三指毒素（3FTxs）代表着弹性毒液中极为多样化的蛋白质家族，从临床角度来看，短链α-神经毒素是最相关的毒素组。本质上，北美珊瑚蛇毒液中3FTxs的变异性和α-神经毒素的比例低，使得很难获得针对主要由这些α-神经毒素引起的毒化症状的有效的抗蛇毒血清。在这项工作中，从墨西哥（M. diastema，M。laticollaris，M。browni和M. tener）的四种珊瑚蛇物种的毒腺中获得了三十个3FTx转录物序列。）。使用基于来自3FTx的高度保守的信号肽的正向寡核苷酸挖掘转录本，这些转录本中的四个名为MlatA1，BD，BE和DH，被编码为短链α-神经毒素。另外，在舒张支原体的毒液中鉴定出一种DHα-神经毒素转录物的同工型。DH转录本中编码的成熟α-神经毒素被异源表达，并且发现它在细菌系统的细胞质中可溶（4.2 mg / l）。重组DH（rD.H）对横纹肌肉瘤细胞（TE671）中表达的肌肉型烟碱型乙酰胆碱受体的IC ₅₀值为31.5±4.4 nM。rDH也有LD ₅₀0.15 mg / kg的小鼠，被评估为新西兰兔的潜在免疫原。测试了兔血清对两种天然珊瑚蛇α-神经毒素和rD.H的保护能力。当在CD1小鼠组中进行测试时，一种抗rD.H兔血清能够中和所有三种神经毒素的致死性。这项工作有助于增进对3FTxs高度多样性的了解，并表明重组珊瑚蛇α-神经毒素是有希望的补充，用于生产抗蛇毒抗蛇毒的超免疫方案。