摘要
选用螯合剂Isothiocyanobenzy-EDTA(ITCBE)络合重金属镉离子,再分别与载体蛋白BSA和KLH相连接,初步制备免疫抗原Cd-ITCBE-KLH和检测抗原Cd-ITCBE-BSA。通过BCA法测定完全抗原浓度,并利用SDS-PAGE电泳、紫外分光光度法及石墨炉原子吸收法对所合成抗原进行初步鉴定;BCA法测出Cd-ITCBE-BSA、Cd-ITCBE-KLH及ITCBE-BSA的实际蛋白浓度依次为1.426,1.504,0.890mg/mL。SDS-PAGE电泳和紫外分光光度法定性的说明抗原合成成功,石墨炉原子吸收法测得完全抗原中镉离子含量最高可达44.56μg/mL,其他无金属抗原、载体蛋白和Hepes中镉离子含量几乎为0,定量地说明了抗原合成成功。
Cadmium ions were coupled to Keyhole Limpet Hemocyanin(KLH) and Bovine Serum Albumin (BSA) via Isothiocyanobenzy-EDTA to gain complete antigen (Cd-ITCBE-KLH and Cd-ITCBE-BSA). The antigen concentration was measured by BCA method; ultraviolet scan of spectrophotometer and SDS- PAGE electmphoresis qualitative identification were done and the content of cadmium ions in antigen was determined by GFAAS method. The actual protein concentration of Cd-ITCBE-BSA, Cd-ITCBE-KLH and 1TCBE-BSA were in the sequence of 1.426,1.504 and 0.890 mg/mL, ultraviolet scan and electmphore- sis identification proved that the antigen synthesis was successful. Concentration of cadmium ion in com- plete antigen was measured as high as 44.56 μg/mL, and that of cadmium ion in ITCBE-BSA, BSA, KIM and Hepes were almost zero, which indicated the hapten was surely coupled into protein, and further tes- tified the antigen synthesis was successful.
出处
《吉林农业大学学报》
CAS
CSCD
北大核心
2013年第2期188-191,共4页
Journal of Jilin Agricultural University
基金
吉林省世行贷款农产品质量安全项目(2011-Z42)
关键词
重金属
镉
人工抗原
heavy metal
cadmium
artificial antigen