抗竹叶青蛇毒IgY抗体：制备、纯化及中和效果
Anti-Trimeresurus albolabris venom IgY antibodies: preparation, purification and neutralization efficacy.
作者信息
Duan Hai-Long, He Qi-Yi, Zhou Bin, Wang Wen-Wen, Li Bo, Zhang Ying-Zheng, Deng Qiu-Ping, Zhang Ying-Feng, Yu Xiao-Dong
机构信息
Animal Toxin Group, Chongqing Key Laboratory of Animal Biology, Chongqing Engineering Research Center of Bioactive Substance, Engineering Research Center of Active Substance and Biotechnology, Ministry of Education, College of Life Science, Chongqing Normal University, Chongqing, 401331 China.
Library, Chongqing Normal University, Chongqing, 401331 China.
出版信息
J Venom Anim Toxins Incl Trop Dis. 2016 Aug 24;22(1):23. doi: 10.1186/s40409-016-0078-3. eCollection 2016.
BACKGROUND
Snakebite incidence in southwestern China is mainly attributed to one of the several venomous snakes found in the country, the white-lipped green pit viper Trimeresurus albolabris. Since antivenom produced from horses may cause numerous clinical side effects, the present study was conducted aiming to develop an alternative antivenom antibody (immunoglobulin Y - IgY) from leghorn chickens.
METHODS
IgY in egg yolk from white leghorn chicken previously injected with T. albolabris venom was extracted by water, precipitated by ammonium sulfate and purified by affinity chromatographic system. IgY was identified by SDS-PAGE, ELISA and Western blot, and its neutralizing assay was conducted on mice.
RESULTS
Chickens injected multiple times with T. albolabris venom elicited strong antibody responses, and from their egg yolk IgY was isolated and purified, which exhibited a single protein band on SDS-PAGE and two bands (about 65 and 35 kDa, respectively) under reduced conditions. Immunoblot analysis revealed that these IgY are polyclonal antibodies since they bind with most venom components. In the neutralizing assay, all mice survived while the ratios of IgY/venom reached up to 3.79 (50.0 mg/13.2 mg).
CONCLUSIONS
IgY antibody response was successfully conducted in white leghorn chicken injected with T. albolabris venom. IgY against T. albolabris venom was obtained for the first time, and it exhibited strong neutralizing potency on mice. These results may lay a foundation for the development of IgY antivenom with clinical applications in the future.
背景
中国西南部的蛇咬伤发生率主要归因于该国发现的几种毒蛇之一，即白唇竹叶青（Trimeresurus albolabris）。由于马源性抗蛇毒血清可能会引起多种临床副作用，因此开展了本研究，旨在从来亨鸡中开发一种替代抗蛇毒抗体（免疫球蛋白Y - IgY）。
方法
从未经处理的来亨鸡中提取卵黄中的IgY，将其注入白唇竹叶青蛇毒中，用水提取，硫酸铵沉淀，并用亲和色谱系统纯化。通过SDS-PAGE、ELISA和Western印迹法鉴定IgY，并在小鼠上进行中和试验。
结果
多次注射白唇竹叶青蛇毒的鸡产生了强烈的抗体反应，并从它们的卵黄中分离和纯化了IgY，其在SDS-PAGE上呈现单一蛋白条带，在还原条件下呈现两条带（分别约为65 kDa和35 kDa）。免疫印迹分析表明，这些IgY是多克隆抗体，因为它们与大多数毒液成分结合。在中和试验中，当IgY/毒液的比例达到3.79（50.0 mg/13.2 mg）时，所有小鼠均存活。
结论
在注射白唇竹叶青蛇毒的来亨鸡中成功诱导了IgY抗体反应。首次获得了针对白唇竹叶青蛇毒的IgY，并且它在小鼠上表现出强大的中和效力。这些结果可能为未来开发具有临床应用价值的IgY抗蛇毒血清奠定基础。
Zotero 插件