应用改良的碳二亚胺法制备无色孔雀石绿完全抗原及鉴定

摘要

The conjugation of paraleucaniline-bovine serum albumin ( PA-BSA) was prepared via a modified carbodiimide method. Features of the conjugation were identified by ultraviolet spectrometry, polyacrylamide gel electrophoresis (SDS-PAGE) and mass spectroscopy. PA-BSA was used as immunogen to immunize Balb/c mice. The results showed that PA-BSA coupling ratio was 37: 1. The titers of purified Pabs were 6. 25×105; Then a competitive inhibition curves of Pabs to LMG was developed, and the IC50 was 11. 8 mg/L. The antibody exhibited high cross-reactivity (100% ) with malachite green and paraleucaniline.%为完善无色孔雀石绿的免疫检测方法,通过改良的碳二亚胺法,改造无色孔雀石绿结构类似物副品红半抗原.经过紫外分光法、十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)、质谱法鉴定,偶联反应成功,且偶联比最高达37∶1.以副品红一牛血清白蛋白偶联物为完全抗原免疫Balb/c小鼠,产生针对无色孔雀石绿的多克隆抗体,效价可达6.25×105；取抗体与无色孔雀石绿进行竞争抑制测定,IC50可以达到11.8 mg/L.该抗体与孔雀石绿及副品红存在交叉反应,交叉反应率均为100％.