Development of a Broad-Spectrum Antiserum against Cobra Venoms Using Recombinant Three-Finger Toxins

• Published in: Toxins Vol. 13; no. 8; p. 556
• Main Authors: Liu, Bing-Sin, Jiang, Bo-Rong, Hu, Kai-Chieh, Liu, Chien-Hsin, Hsieh, Wen-Chin, Lin, Min-Han, Sung, Wang-Chou
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 10.08.2021; MDPI
• Subjects: Animals; Antibodies; Antibodies, Neutralizing - blood; Antibody response; Antigens; Antisera; antiserum; Antivenins - administration & dosage; Antivenom; broad spectrum; Chemical bonds; Chromatography; Circular dichroism; cobra; Cytotoxicity; Dichroism; Disulfide bonds; E coli; Elapid Venoms - immunology; Elapid Venoms - toxicity; Elapidae; Escherichia coli - genetics; Fusion protein; Horses; Immunization; Lethality; Low temperature; Mass spectrometry; Mass spectroscopy; Mice; Mice, Inbred ICR; Naja; Neurotoxicity; Neurotoxins; Neurotoxins - immunology; Neurotoxins - toxicity; Neutralization; Peptides; Protein disulfide-isomerase; Proteins; Rabbits; recombinant protein; Recombinant Proteins - administration & dosage; Recombinant Proteins - chemistry; Scientific imaging; Snake bites; Snakes; three finger toxins; Three Finger Toxins - administration & dosage; Three Finger Toxins - chemistry; Three Finger Toxins - genetics; Toxicity; Toxins; Venom; Asia; cobra; three finger toxins; antiserum; immunization; recombinant protein; broad spectrum
• ISSN: 2072-6651; 2072-6651
• DOI: 10.3390/toxins13080556

Three-finger toxins (3FTXs) are the most clinically relevant components in cobra (genus ) venoms. Administration of the antivenom is the recommended treatment for the snakebite envenomings, while the efficacy to cross-neutralize the different cobra species is typically limited, which is presumably due to intra-specific variation of the 3FTXs composition in cobra venoms. Targeting the clinically relevant venom components has been considered as an important factor for novel antivenom design. Here, we used the recombinant type of long-chain α-neurotoxins (P01391), short-chain α-neurotoxins (P60770), and cardiotoxin A3 (P60301) to generate a new immunogen formulation and investigated the potency of the resulting antiserum against the venom lethality of three medially important cobras in Asia, including the Thai monocled cobra ( ), the Taiwan cobra ( ), and the Thai spitting cobra ( ) snake species. With the fusion of protein disulfide isomerase and the low-temperature settings, the correct disulfide bonds were built on these recombinant 3FTXs (r3FTXs), which were confirmed by the circular dichroism spectra and tandem mass spectrometry. Immunization with r3FTX was able to induce the specific antibody response to the native 3FTXs in cobra venoms. Furthermore, the horse and rabbit antiserum raised by the r3FTX mixture is able to neutralize the venom lethality of the selected three medically important cobras. Thus, the study demonstrated that the r3FTXs are potential immunogens in the development of novel antivenom with broad neutralization activity for the therapeutic treatment of victims involving cobra snakes in countries.