两种方法制备氟苯尼考人工抗原及其鉴定

目的：通过两种不同方法制备氟苯尼考人工抗原并将制备的人工免疫原用于动物免疫。方法：分别采用琥珀酸酐法和强碱水解法制备两种人工半抗原(HP1 和HP2)。将两种半抗原与牛血清白蛋白(BSA)和卵清白蛋白(OVA)通过活泼酯法和戊二醛法进行偶联，制备人工免疫原和人工包被原，分别命名为HP1-BSA、HP1-OVA、HP2-BSA 和HP2-OVA。分别将两种方法合成的人工免疫原用于动物免疫，用两种人工包被原对获得的两种抗血清进行评价。结果：经质谱、紫外光谱表征，氟苯尼考两种半抗原均合成成功，并且与载体蛋白的偶联比分别为12.4:1(HP1-BSA)、8.5:1(HP1-OVA)、21.8:1(HP2-BSA)、15.4:1(HP2-OVA)。动物实验结果表明，获得的针对HP1-BSA的抗血清与HP2-OVA 包被原结合最为灵敏。结论：活泼酯法和戊二醛法均成功制备了氟苯尼考人工抗原。

Preparation of Haptens by Two Different Methods for Immunoassay Determination of Florfenicol and Their Identification

Two hapaten synthesis methods, succinic anhydride method and alkaline hydrolysis were used for florfenicol deriviazation and the synthesized hapatens, named HP1 and HP2, were separately conjugated with BAS (bovine serum albumin) by active ester method and OVA (ovalbumin) by glutaraldehyde method to obtain two immunoagents, named HP1-BAS and HP2-BAS and two coating antigens, named HP1-OVA and HP2-OVA. HP1-BAS and HP2-BAS were separately used to immunize Balb/c mice for obtaining antisera. Antisera response to HP1-BAS and HP2-BAS were evaluated for their titers using HP1-OVA and HP2-OVA. The mass spectrometric and UV spectral characterization demonstrated that HP1 and HP2 were successfully synthesized. The coupling ratios between HP1 or HP2 and BAS or OVA were determined to be 12.4:1 (HP1-BSA), 8.5:1 (HP1-OVA), 21.8:1 (HP2-BSA) and 15.4:1 (HP2-OVA). The antiserum responses to HP1-BAS had the most specificity to HP2-OVA.