抗蓖麻毒素精制免疫球蛋白的制备及免疫防治效果评价

Preparation of purified immunoglobulin against ricin and evaluation of its immune control effect

研制抗蓖麻毒素精制免疫球蛋白,即马抗蓖麻毒素免疫球蛋白F(ab′)_2制品,为蓖麻毒素中毒的防治提供特效药。利用传统方法,从蓖麻籽中提取、纯化脱毒蓖麻毒素,加入适量弗氏佐剂充分乳化,免疫马匹,测定免疫马匹血清中和抗体效价并采血,分离血清,按照免疫球蛋白F(ab′)_2生产工艺制备治疗性抗体成品,并对成品进行中和抗体效力检测、安全性检测及免疫防治效果评价。结果显示,纯化的蓖麻毒素抗原,经HPLC法测定纯度大于90%,腹腔注射对小鼠的半数致死量(LD_(50))约为6.38μg/kg;免疫马匹血清中和抗体效价达1∶64000以上时,采血并制备F(ab′)_2成品,纯度在80%以上,中和抗体效价均超过6000 U/mL;安全性符合现行《中国药典》要求,对蓖麻毒素中毒动物模型有很好的免疫防治效果。本研究建立了蓖麻毒素精制免疫球蛋白制品生产工艺,制备的成品符合现行《中国药典》要求,为蓖麻毒素中毒的防治提供了新的救命药。

The purpose of this study is to develop the purified immunoglobulin against ricin and provide a specific drug for the prevention and treatment of ricin poisoning.The traditional method was used to extract,purify and detoxify ricin from castor seeds,then the ricin was emulsified in Freund’s adjuvant and used to immunize horses.The serum neutralizing antibody titer of immunized horses was determined and then blood was collected to separate serum to prepare therapeutic antibody products according to the production process of immunoglobulin F(ab′)2.Neutralizing potency of the products was tested,the safety and immune control effect were also evaluated.The results showed that the purity of purified ricin antigen was more than 90%,and the median lethal dose of intraperitoneal injection was about 6.38μg/kg.When the neutralizing antibody titer of immunized horse serum was more than 1∶64000,the F(ab′)2was prepared from the blood,its purity was more than 80%,and the neutralizing antibody titer was more than 6000U/mL.The safety of the product meets the requirements of current Chinese Pharmacopoeia and has good immune effect on animal model of ricin toxicosis.This study provides a new life-saving drug for the prevention and treatment of ricin poisoning.