title: Development of ELISA for detection of mercury based on specific monoclonal antibodies against mercury-chelate.
abstract: Immunoassays for heavy metals offer an alternative approach to traditional techniques for detection of mercury. In this study, a mercury-chelate was prepared with 1-(4-aminobenzyl) ethylenediamine-N,N,N',N'-tetraacetic acid (aminobenzyl-EDTA). The resulting complex was linked to keyhole limpet hemocyanin (KLH) or bovine serum albumin via the amino group and used as the immunizing antigen or detection antigen, respectively. BALB/c mice were immunized with KLH-aminobenzyl-EDTA-Hg and spleen cells from BALB/C mice were fused with Sp2/0 cells. One cell line (5F7) produced monoclonal antibodies with preferential selectivity and sensitivity for aminobenzyl-EDTA-Hg. This cell line had an affinity constant of 4.31 × 10(9) L/mol and its cross-reactivity (CR) with other metals was <2%. The antibody was used for competitive indirect ELISA (CI-ELISA) for Hg(2+) measurements. The detection range was 0.087-790.4 μg/L and the lower limit of detection was 0.042 μg/L. The concentrations of mercury in environmental water samples obtained by CI-ELISA correlated well with graphite furnace atomic absorption spectrometry (GFAAS), and the mean recovery was 88.82% to 104.64%. These results indicate that this method could be used for monitoring mercury of water.
authors: College of Animal Science and Veterinary Medicine, Jilin University, 5333 Xi'an Road, Changchun, 130062, China.
authors: Yi
authors: Y
authors: Zhang
authors: Xiaobing
authors: X
authors: Li
authors: Guowen
authors: G
authors: Liu
authors: Zhe
authors: Z
authors: Wang
authors: Tao
authors: T
authors: Kong
authors: Jiajia
authors: J
authors: Tang
authors: Peng
authors: P
authors: Zhag
authors: Wei
authors: W
authors: Yang
authors: Dongna
authors: D
authors: Li
authors: Lei
authors: L
authors: Liu
authors: Guanghong
authors: G
authors: Xie
authors: Jianguo
authors: J
authors: Wang
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000302:isolation & purification; D000915:Antibody Affinity; D000918:Antibody Specificity; D045744:Cell Line, Tumor; D002614:Chelating Agents / Q000737:chemistry*; D002681:China; D003429:Cross Reactions; D004492:Edetic Acid / Q000031:analogs & derivatives / Q000737:chemistry; D004784:Environmental Monitoring / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D006433:Hemocyanins / Q000737:chemistry; D006825:Hybridomas; D007202:Indicators and Reagents; D008628:Mercury / Q000032:analysis* / Q000276:immunology; D008670:Metals / Q000032:analysis; D051379:Mice; D008807:Mice, Inbred BALB C; D012015:Reference Standards; D045483:Rivers; D012710:Serum Albumin, Bovine / Q000737:chemistry; D014874:Water Pollutants, Chemical / Q000032:analysis
chemical_list: D000911:Antibodies, Monoclonal; D002614:Chelating Agents; D007202:Indicators and Reagents; D008670:Metals; D014874:Water Pollutants, Chemical; D012710:Serum Albumin, Bovine; C057690:aminobenzyl-EDTA; D006433:Hemocyanins; D004492:Edetic Acid; C032808:keyhole-limpet hemocyanin; D008628:Mercury