摘要
目的 为螨性疫苗及其原材料标准化的检测、质量的监控以及进一步开发提供相应的技术。方法 应用层析介质以及蛋白沉淀技术从螨代谢培养基中分离数种重要的以及国际公认的螨性分泌排泄致敏蛋白Dff F1、Dff F2、Der fⅡ和Der fⅢ,经临床检验和放射性变应原吸附试验(RAST)等鉴定其免疫原性后,分别制备抗血清,再结合免疫学试验对疫苗制剂进行抗原的标准化分析。结果 4种成分除Dff F1外,均呈较强的变应原性,且与哮喘病人血清IgE显示出高结合率;火箭电泳峰清晰明了,ELISA的A值与对照组比较差异有显著性。结论 纯化的Dff F1、Dff F2、Der fⅡ和Der fⅢ致敏蛋白多克隆抗体结合免疫相关技术完全可以用于检测候选螨性疫苗口服液制剂中抗原的性质、含量等,以及进行标准化分析。
Objective To serve as an useful tool for standardization of allergens and survey of quality of the oral vaccine against Dermatophagoides farinae and develop new products. Methods Several allergenic proteins(Dff F1, Dff F2, Der f Ⅱ and Der fⅢ ) from the metabolic medium of D. farinae were purified by chromatography and sedimentation techniques, and identified by clinical test, Radioallergosorbent test(RAST) and some immuno-tests. Results All fractions except Dff F1 were showed very strongly al-lergic, and recognized highly with IgE from sera of patients with asthma. Polyclonal antibodies anti-Dff F1, Dff F2, Der fⅡand Der f Ⅲ were purified from sera of the immunized rabbits and used for detection of the major components, proportion of allergic proteins, and standardized the antigens of oral vaccines against dust mite. The peaks of Rocket Electrophoresis look clear, and the A values of ELISA were significant differences between the test groups and controls. Conclusion These purified polyclonal antibodies against Dff F1, Dff F2, Der fⅡ and Der fⅢwith immuno-techniques can be used for standardization of allergen of the oral vaccine.
出处
《中国寄生虫病防治杂志》
CSCD
2003年第4期224-228,共5页
Chinese Journal of Parasitic Disease Control
