Zhu Haiting, Pan Yuexin, Tai Zhiyuan, Wang Mingqian, Liu Xia, Yu Xiaodong, He Qiyi
Engineering Research Center of Active Substance and Biotechnology, Ministry of Education, College of Life Science, Chongqing Normal University, Chongqing, China.
J Venom Anim Toxins Incl Trop Dis. 2025 May 9;31:e20240060. doi: 10.1590/1678-9199-JVATITD-2024-0060. eCollection 2025.
, or the hundred-pace snake, poses severe health risks due to its venom. Envenomation by this snake leads to complications such as hemorrhage, edema, and coagulopathy. Traditional antivenoms are limited by venom variability and often contain non-neutralizing antibodies, highlighting the need for more precise and effective immunogens.
This study utilized epitope-based antibody technology to develop a targeted sera against venom metalloproteinases (MPs) and phospholipases A (PLAs). Twelve antigenic epitopes were identified via bioinformatics, leading to the design of a composite antigen peptide, EpiMPLA. It was engineered to be expressed via two expression systems, resulting in the recombinant immunogens, ProMPLA and p2AMPLA.
Immunization with ProMPLA and p2AMPLA produced robust antibody responses in mice, effectively inhibiting MPs and PLAs. assays demonstrated that sera from immunized mice reduced the activity of these venom enzymes, minimized venom-induced hemorrhage and edema, and restored blood coagulation. At a venom dose of 2×LD, all mice in the control group died, while survival rates were 90% for anti-ProMPLA and 70% for anti-p2AMPLA.
The EpiMPLA epitope represents a promising candidate for generating neutralizing antibodies against venom, demonstrating its potential to address critical gaps in current antivenom therapy. These findings not only validate the feasibility of epitope-based antivenom development but also pave the way for further research to optimize this strategy.
五步蛇，因其毒液会带来严重的健康风险。被这种蛇咬伤会导致出血、水肿和凝血病等并发症。传统抗蛇毒血清受毒液变异性限制，且常含有非中和抗体，这凸显了对更精确有效免疫原的需求。
本研究利用基于表位的抗体技术开发针对毒液金属蛋白酶（MPs）和磷脂酶A（PLAs）的靶向血清。通过生物信息学鉴定出12个抗原表位，从而设计出复合抗原肽EpiMPLA。它经工程改造可通过两种表达系统表达，产生重组免疫原ProMPLA和p2AMPLA。
用ProMPLA和p2AMPLA免疫小鼠产生了强烈的抗体反应，有效抑制了MPs和PLAs。实验表明，免疫小鼠的血清降低了这些毒液酶的活性，将毒液诱导的出血和水肿降至最低，并恢复了血液凝固。在毒液剂量为2×LD时，对照组所有小鼠死亡，而抗ProMPLA组的存活率为90%，抗p2AMPLA组为70%。
EpiMPLA表位是产生抗毒液中和抗体的一个有前景的候选者，证明了其在解决当前抗蛇毒血清疗法关键差距方面的潜力。这些发现不仅验证了基于表位的抗蛇毒血清开发的可行性，也为进一步优化该策略的研究铺平了道路。
