重金属镉离子人工抗原的合成与鉴定

Synthesis and Identification of Artificial Antigen for Heavy Metal Cadmium Ions

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.