鼠疫组分疫苗F1抗原、rV抗原含量的检测及其质量控制

目的采用双抗体夹心ELISA法检测鼠疫组分疫苗中F1和rV的抗原含量。方法利用F1、rV抗原单克隆抗体,对鼠疫组分疫苗原液进行抗原鉴别试验;分别应用F1、rV抗原双抗体夹心ELISA法对疫苗原液进行抗原定量检测;验证A(l OH)3佐剂吸附抗原的完全性;将疫苗成品于4℃放置18个月,分别于1和18个月时取样,用建立的双抗体夹心ELISA法检测F1和rV抗原含量,分析抗原的稳定性;对3批鼠疫菌rV抗原原液3步纯化工艺进行验证。结果鼠疫组分疫苗原液中的F1和rV抗原具有良好的反应原性;用建立的双抗体夹心ELISA法检测4批F1和rV抗原原液中F1和rV抗原的含量与Lowry法检测结果的误差均在±20%以内;4批鼠疫组分疫苗离心后的上清液中均未检测到F1和rV抗原,表明A(lOH)3佐剂吸附抗原完全;4℃保存18个月,疫苗中的F1和rV抗原含量稳定;3批鼠疫菌rV抗原原液经阴离子交换层析、疏水层析、凝胶过滤层析3步纯化,rV抗原在纯度增加的同时,抗原含量也增加。结论已建立的F1、rV抗原双抗体夹心ELISA法可用于鼠疫组分疫苗中F1和rV抗原的定量检测。

Determination of F1 and rV antigen contents and quality control of plague component vaccine

Objective To determine the F1 and rV antigen contents in plague component vaccine by double antibody sandwich ELISA.Methods The antigens in bulk of plague component vaccine were identified with monoclonal antibodies(McAbs)against F1 rV antigens,and determined quantitatively by double antibody sandwich ELISA.The completeness of antigen adsorbed onto aluminium hydroxide adjuvant was verified.The final product of vaccine was stored at 4 ℃ for 18 months,from which samples were taken 1 and 18 months after storage respectively and determined for F1 and rV antigen contents by the developed double antibody sandwich ELISA method to analyze the stability of antigen.The three-step purification procedure for three batches of bulk rV antigen of Yersiria pestis were verified.Results The F1 and rV antigens in bulk of plague component vaccine showed good reactogenicity.The F1 and rV antigen contents in four batches of bulks were determined by the developed method,of which the results showed an error of not more than 20% with those determined by Lowry method.No F1 or rV antigen was detected in the centrifuge supernatant of four batches vaccine,indicating that the antigens were completely adsorbed by aluminium hydroxide adjuvant.The F1 and rV antigen contents in vaccine were stable after storage at 4 ℃ for 18 months.After three batches of bulk were purified by anion exchange,hydrophobic and gel filtration chromatography,both the purity and content of rV antigen increased.Conclusion The developed double antibody sandwich ELISA method may be used for the quantitative determination of F1 and rV antigens in plague component vaccine.