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目的:ELISA法检测双黄连注射剂中小分子半抗原物质绿原酸致敏家兔血清中的特异性抗体效价。方法:取新西兰种家兔,自家兔耳中心动脉采血10ml,制备致敏前血清。将绿原酸与家兔血清体外孵育,制成绿原酸-血浆蛋白完全抗原,将完全抗原皮下注射以免疫动物,1月内共5次。末次注射后7天,麻醉动物,自腹主动脉取血,制备血清,即为绿原酸-血浆蛋白全抗原的抗血清,用ELISA法检测抗血清的抗体效价(OD值);间接竞争ELISA法测定家兔抗血清的特异性抗体;采用兔免疫球蛋白E(Ig E)ELISA试剂盒检测家兔抗血清中Ig E抗体的含量。结果:绿原酸的抗体效价高于致敏前血清OD值的2倍,说明绿原酸有潜在的致敏性。绿原酸免疫后的家兔抗血清中的抗体具有一定的特异性。经过多次免疫后的家兔体内产生了外源性Ig E型抗体。结论:绿原酸在本实验条件下对家兔有致敏性。
Objective: To detect the titer of specific antibody in sera of mice sensitized with chlorogenic acid, a small molecule hapten substance, of Shuanghuanglian injection. Methods: New Zealand rabbits were taken from the central ear artery of rabbits and 10ml blood samples were collected to prepare pre-sensitized serum. Chlorogenic acid and rabbit serum in vitro incubation, made of chlorogenic acid - plasma protein complete antigen, subcutaneous injection of complete antigen to immunize animals, a total of 5 times a month. Seven days after the last injection, the animals were anesthetized and blood was taken from the abdominal aorta to prepare serum, which is the antiserum of chlorogenic acid-plasma protein antigens. The antibody titer (OD) of the antisera was measured by ELISA. Indirect competitive ELISA Method to determine the specific antibody of rabbit antiserum. The content of Ig E antibody in rabbit antiserum was detected by rabbit immunoglobulin E (Ig E) ELISA kit. RESULTS: The antibody titer of chlorogenic acid was 2 times higher than the OD value of serum before priming, indicating that chlorogenic acid has potential allergenicity. Antibodies in rabbit antisera after immunization with chlorogenic acid have certain specificity. After repeated immunization of rabbits in vivo produced exogenous Ig E-type antibodies. Conclusion: Chlorogenic acid is sensitized to rabbits in this experiment.