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基于IgY(DeltaFc)抗体的神经毒素抗蛇毒血清的研制及其中和效力研究。

The development of IgY(DeltaFc) antibody based neuro toxin antivenoms and the study on their neutralization efficacies.

作者信息

Chiou Victor Y-Neng

机构信息

Good Biotech Corp., Taichung, Taiwan, Republic of China.

出版信息

Clin Toxicol (Phila). 2008 Jul;46(6):539-44. doi: 10.1080/15563650701771973.

Abstract

INTRODUCTION

Immunotherapy for treatment of snake bites has been based on mammalian IgG. Recently, polyvalent ovine Fab has become available. However, papain, used in the Fab fragmentation process, is a human allergen. Avian eggs are a source of antibodies and a truncated version of IgY, IgY(DeltaFc), is found in ducks. In this study, we induced duck antibodies by using detoxified cobra and krait venoms and then purified IgY(DeltaFc) antibodies from the hyperimmune duck egg yolk.

METHODS

Ducks were used for immunization and their eggs were collected for antibody production. ICR strain female mice were used in the in vivo neutralization test. Monovalent antivenoms to Formosan cobra venom and Formosan multi-banded krait venom were raised and purified from hyper-immune duck egg yolk individually. The LD(50) of venoms were determined by subcutaneous injection of different venom doses into the mice. The survival/death ratios were recorded after 24 hours.

RESULTS

The antibody purified from egg yolk showed high titer response to its immunogen (cobra or krait venom) by an ELISA. Overall, the antibodies from duck eggs efficiently protected mice from envenomations.

DISCUSSION

The antivenoms purified from the egg yolk of ducks immunized with cobra venom and krait venom neutralized the lethal effects of these venoms with good efficacy in a mouse model. The antivenoms were effective in neutralizing lethality in mice injected at 4xLD(50) of venoms.

CONCLUSIONS

These results indicate that antibodies derived from ducks can serve as a new source for the generation of antivenoms.

摘要

引言

用于治疗蛇咬伤的免疫疗法一直基于哺乳动物IgG。最近,多价羊Fab片段已可获得。然而,Fab片段化过程中使用的木瓜蛋白酶是一种人类过敏原。禽蛋是抗体的来源,鸭体内存在IgY的截短版本IgY(DeltaFc)。在本研究中,我们使用解毒的眼镜蛇毒和银环蛇毒诱导鸭产生抗体,然后从超免疫鸭蛋黄中纯化IgY(DeltaFc)抗体。

方法

使用鸭进行免疫,并收集其蛋用于抗体生产。在体内中和试验中使用ICR品系雌性小鼠。分别从超免疫鸭蛋黄中制备并纯化针对台湾眼镜蛇毒和台湾多带银环蛇毒的单价抗蛇毒血清。通过向小鼠皮下注射不同剂量的毒液来测定毒液的半数致死量(LD(50))。24小时后记录存活/死亡比率。

结果

通过ELISA检测,从蛋黄中纯化的抗体对其免疫原(眼镜蛇或银环蛇毒)显示出高滴度反应。总体而言,鸭蛋黄中的抗体有效地保护小鼠免受蛇毒侵害。

讨论

用眼镜蛇毒和银环蛇毒免疫的鸭蛋黄中纯化的抗蛇毒血清在小鼠模型中能有效中和这些毒液的致死作用。这些抗蛇毒血清对注射4倍LD(50)毒液的小鼠的致死性具有中和作用。

结论

这些结果表明,鸭源抗体可作为产生抗蛇毒血清的新来源。

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