展青霉素多克隆抗体的制备

Preparation of Polyclonal Antibodies for Patulin

展青霉素是一种神经毒素,对动物的细胞和组织有很强的毒性作用。展青霉素原药毒性大,价格昂贵且不稳定;其化学结构简单,直接制备免疫原效果不理想。因此,目前报道的展青霉素免疫分析方法并不多。免疫原的设计合成是免疫分析方法建立的关键步骤之一。本论文通过计算机辅助模拟设计,借鉴药物分子虚拟筛选的方法,研究展青霉素结构特点,优化筛选毒性较低且价格廉价的替代物,设计并成功合成展青霉素半抗原。活化酯法偶联两种载体蛋白(BSA和KLH)制备展青霉素两种免疫原,免疫动物制备多克隆抗体,抗血清效价分别可以达到1:16000和1:64000。本研究为进一步建立展青霉素免疫分析方法打下基础。同时,论文为小分子有毒有害物免疫原设计提供了一种新思路和新方法。

Patulin is a neurotoxin with a strong toxic effect. This toxin is expensive and unstable; the chemical structure of patulin is simple, and the direct preparation of immunogens from patulin has a poor immune effect. As a result, few immunoassay methods for patulin have been reported. Design and preparation of the immunogen is one of the key steps for developing the immunoassay. Here, the structural characteristics of patulin were studied via computer-aided simulation design and methods for virtual screening of drug molecules. A suitable substitute with low toxicity and cost was optimized and screened, and the hapten of patulin was designed and successfully prepared. Two immunogens of patulin were obtained by conjugation with two carrier proteins （BSA and KLH） by the active ester method. The polyclonal antibodies were prepared aiter the immunization of rabbits, and the antiserum titers reached 1：16000 and 1：64000, respectively. This study lays a foundation for the further establishment of an immunoassay for patulin and also provides new ideas and methods for designing immunogens for toxic small-molecule compounds.