Lei Hongyu, Yuan Hui, Wu Jing, Yuan Liyun, Wen Lixin, Ni Hengjia
College of Veterinary Medicine, Hunan Agricultural University, Changsha 410128, China.
Sheng Wu Gong Cheng Xue Bao. 2008 May;24(5):898-902.
To establish a new immune assay for Penicillic Acid (PA) from Penicillium cyclopium, we studied the synthesis of conjugated complete antigens for penicillic acid. PA was conjugated to bovine serum album (BSA) and ovalbumin (OVA) by 1-ethyl-3-(3-dimethyl-aminopropyl) carbodiimide hydrochloride (EDC). The artificial antigens PA-BSA and PA-OVA were identified by ultraviolet spectrometric scanning, SDS-PAGE and immunization. Results showed that the absorption peak of conjugation were different from that of the carrier protein alone and of the PA. The conjugated ratio of PA and BSA was 23.2:1 and that of PA and OVA was 10.4:1. Balb/c mice were immunized by the artificial antigen of PA-BSA, with PA-OVA as coating antigen. The average titer of antiserums was more than 12 800 by indirect ELISA. The obtained antigens offered a basis for developing immunoassay method.
为建立一种针对环孢青霉中青霉素酸(PA)的新型免疫检测方法,我们研究了青霉素酸偶联完全抗原的合成。通过1-乙基-3-(3-二甲基氨基丙基)碳二亚胺盐酸盐(EDC)将PA与牛血清白蛋白(BSA)和卵清蛋白(OVA)偶联。通过紫外光谱扫描、SDS-PAGE和免疫实验对人工抗原PA-BSA和PA-OVA进行鉴定。结果表明,偶联物的吸收峰与单独的载体蛋白以及PA的吸收峰不同。PA与BSA的偶联比为23.2:1,PA与OVA的偶联比为10.4:1。以PA-OVA作为包被抗原,用PA-BSA人工抗原免疫Balb/c小鼠。通过间接ELISA法测得抗血清的平均效价超过12800。所获得的抗原为开发免疫检测方法提供了基础。