【摘要】 目的:制备重金属Cr3+的单克隆抗体(mAb)并建立间接竞争ELISA法,用于Cr3+的检测。方法:用合成的重金属铬-螯合剂-BSA抗原免疫BALB/c小鼠,通过细胞融合、克隆化和ELISA等技术建立杂交瘤细胞株;腹水型mAb经硫酸铵沉淀纯化后建立间接竞争ELISA法并与9种金属进行交叉实验;水样加标后,进行间接竞争ELISA法与ICP-AES法检测Cr3+的比较,计算两种方法符合率。结果:获得3株稳定分泌抗重金属Cr3+mAb的杂交瘤细胞株,其中5G12C5株mAb间接ELISA效价>1∶1×105,间接竞争ELISA法检测Cr3+的范围为0.783~50μg/L,最低检测限为1μg/L,与Fe3+存在<10%交叉,而与其他金属无交叉反应;当水样中加标量>1μg/L时,间接竞争ELISA法与ICP-AES法检测Cr3+的符合率达96.95%。结论:获得效价高、特异性强的重金属Cr3+mAb,初步建立了检测Cr3+的间接竞争ELISA法,该方法检测的灵敏度可达到国家地下Ⅰ类水的检测标准(0.005 mg/L)。更多还原

【Abstract】 AIM:To produce the monoclonal antibodies(mAb) against Heavy Metal Chromium,and develop a competitive inhibition enzyme-linked immunosorbent assay(ciELISA) for the detection of Chromium.METHODS: Chromium ions was chelated with bifunctional chelating agent Isothio-cyanobenzyl-EDTA firstly and then conjugated with bovin serum alburmin(BSA) and ovalbmin(OVA) respectively to complete antigens.Four female BALB/c mice were immunized with Cr-iEDTA-BSA,the Hybridoma lines secreting monoclonal antibodies against Chromium ions were established by the hybridoma technology.The titer and the specificity of the antibodies were characterized in the way of indirect enzym-elinked immunosorbent assay(iELISA) and ciELISA.RESULTS: After immunization and cell fusion,three hybridomaes which can stably secrete mAbs against chromium were obtained.The antibody titer was up to 1×105,only had 9.62% cross-reactivity to Fe3+ and little to other ions.Linear detection of ciELISA for Chromium covered a range from 0.783 μg/L-50 μg/L with the limit at the level of 1 μg/L.An excellent correlation of results obtained by ciELISA and ICP-AES when the concentration more than 1 μg/L.CONCLUSION: The monoclonal antibody against Chromium with high sensitivity and specificity has been generated and a competitive inhibition enzyme-linked immunosorbent assay for the detection of Chromium ions has been established successfully,which was met the the national standard of water quality nonitoring.更多还原