摘要
目的建立并初步验证吸附DTaP-脊髓灰质炎灭活疫苗(inactivated poliovirus vaccine,IPV)(DTaP-IPV)的D抗原检测预处理解吸附方法.方法选用4种不同解吸附剂,在相同条件下对DTaP-IPV进行解吸附,用间接ELISA检测IPV D抗原.根据各型D抗原的检测结果,确定最适解吸附剂和解吸附条件,并对建立的方法进行初步验证.结果对分别用4种解吸附剂处理的同批疫苗检测D抗原显示,乙二胺四乙酸二钠(ethylenediamine tetraacetic acid disodium,EDTA-2Na)解吸附剂和200 g/L柠檬酸三钠解吸附剂的解离效果好于另2种解吸附剂.进一步对解离效果较好的2种解吸附剂进行作用时间和温度研究显示,EDTA-2Na解吸附剂于25℃处理16~20 h的解离效果最好,IPVⅠ、Ⅱ、Ⅲ型D抗原的回收率分别为102%、101%、103%.建立的方法的重复性和中间精密度良好,经该法解吸附的DTaP-IPV的Ⅰ、Ⅱ、Ⅲ型D抗原检测结果的变异系数均小于10%.结论建立的解吸附方法能有效解吸附DTaP-IPV,可用于DTaP-IPV D抗原检测预处理.
Objective To establish and preliminarily validate a desorption method for detection pretreatment of D antigens in adsorbed DTaP-inactivated poliovirus vaccine (IPV)(DTaP-IPV). Methods Four kinds of desorbents were selected to desorb DTaP-IPV under same conditions. IPV D antigens were detected by indirect ELISA. According to the detection results of different types of D antigens, the optimum desorbent and desorption condition were determined. The established method was validated preliminarily. Results D antigens in the same batch of DTaP-IPV desorbed with 4 kinds of desorbents respectively were detected. The results showed that desorption effect of ethylenediamine tetraacetic acid disodium (EDTA-2Na) desorbent and 200 g/L trisodium citrate desorbent were better than that of other two desorbents. Two desorbents with better desorption effect were further studied on treatment time and temperature. The results showed that the desorption effect of EDTA-2Na desorbent treating at 25 ℃ for 16-20 h was the best. The average recovery rates of IPV type Ⅰ,Ⅱ,Ⅲ D antigens were 102%, 101% and 103%, respectively. The established method had good repeatability and intermediate precision. The variation coefficients of detection results of type Ⅰ,Ⅱ,Ⅲ D antigens in DTaP-IPV desorbed by the established method were all less than 10%. Conclusion The established desorption method can effectively desorb DTaP-IPV, and can be used for detection pretreatment of D antigens in DTaP-IPV.
作者
殷建齐
尹珊珊
邓海清
张弦弦
邢盛宇
顾行举
任玉莹
韩菲
宋琳琳
季庚辰
刘建凯
Yin Jianqi;Yin Shanshan;Deng Haiqing;Zhang Xianxian;Xing Shengyu;Gu Xingju;Ren Yuying;Han Fei;Song Linlin;Ji Gengchen;Liu Jiankai(R & D Center, Beijing Minhai Biotechnology Co., Ltd., Beijing 102600, China)
出处
《国际生物制品学杂志》
CAS
2019年第4期171-174,共4页
International Journal of Biologicals
