苊人工抗原制备及两种间隔臂长对抗原免疫效果的影响

Preparation of artificial antigens for acenaphthene and the influences of two kinds of spacer arm lengths on immunizing effects of the antigens

多环芳烃是一类含有两个以上苯环的碳氢化合物,广泛存在于大气、水体、土壤、食品中,苊是多环芳烃的一种。为了制备免疫效果好的完全抗原从而为免疫学检测方法建立基础,分别以3-苊丁酸和5-苊甲酸与牛血清白蛋白(BSA)或卵清蛋白(OVA)进行偶联,采用活泼酯法成功制备了3-苊丁酸-BSA、5-苊甲酸-BSA两种间隔臂长的免疫用人工抗原及3-苊丁酸-OVA检测用人工抗原,并通过非变性聚丙烯酰胺凝胶电泳和紫外扫描法对人工抗原进行了鉴定。分别用两种免疫抗原足垫免疫Balb/c小鼠,通过测定免疫小鼠血清抗体的效价和特异性,分析不同间隔臂长免疫抗原的免疫效果。结果表明:4碳臂(3-苊丁酸-BSA)较1碳臂(5-苊甲酸-BSA)产生了更高苊的特异性抗体。

Polycyclic aromatic hydrocarbons （PAHs） are a class of carcinogenic hydrocarbons containing two or more benzene rings, widely present in the atmosphere, water, soil, food. Acenaphthene is a kind of PAHs. To prepare a complete antigen with good immune effect and lay a foundation for immunological detection method, acenaphthene - 3 - butyric acid and acenaphthene - 5 - methane acid were conjugated to bo- vine serum albumin （BSA） or ovalbumin （ OVA）, respectively. Artificial antigens of acenaphthene -3 - butyric acid - BSA and acenaphthene - 5 - methane acid - BSA with two kinds of spacer ann lengths for immunization and the artificial antigen for the detection of acenaphthene - 3 - butyric acid - BSA were successfully prepared using an active ester method. The artificial antigens were identified by non - denaturing poly- acrylamide coagulation gel electrophoresis and an ultraviolet scanning method. Two kinds of immunogen footpads were used to immunize Balb/ c mice, and then the immunizing effects of immunogen with different spacer arm lengths were analyzed through the determination of serum anti- body titer and specificity of immunized mice. The result showed that 4 - carbon spacer arm immunogen （ acenaphthene - 3 - butyric acid - BSA） could produce a higher specific antibody of acenaphthene compared with one carbon spacer ann immunogen （ acenaphthene -5 - methane acid - BSA）.