title: Preparation of novel monoclonal antibodies against chelated cadmium ions.
abstract: The detection of cadmium ions using enzyme-linked immunosorbent assays (ELISA) has been reported by several research groups. Because cadmium ions are too small to stimulate the immune system, high molecular weight immunogens of cadmium are constructed using bifunctional chelators. At present, the most commonly used bifunctional chelator for the preparation of antigens for heavy metal ions is 1-(4-isothiocyanobenzyl) ethylenediamine N,N,N',N'-tetraacetic acid (ITCBE). However, the price of ITCBE is high. So we are interested in a cheaper bifunctional chelator, 1-(4-aminobenzyl) ethylenediamine N,N,N',N'-tetraacetic acid (aminobenzyl-EDTA). Here, cadmium ions were conjugated to carrier proteins using aminobenzyl-EDTA to make artificial antigens. Then, several mice were immunized with the antigen. And monoclonal antibodies (MAbs) against cadmium were produced. Spleen cells of immunized mice were fused with myeloma cells. The resulting hybridomas were screened using protein conjugates which were covalently bound to metal-free EDTA or cadmium. Three hybridoma cell lines (A3, E4 and B5) that produced MAbs with high selectivity and sensitivity were expanded for further study. Cross-reactivities with other metals were below 1 %. These antibodies were used to construct competitive ELISAs. The IC50 for A3 was 8.4 μg/l. The detection range and the lowest detection limit using the antibody A3 was 0.394-64.39 and 0.051 μg/l, respectively. Spike-recovery studies in tap water showed that the antibody A3 could be used for cadmium detection in drinking water.
authors: College of Animal Science and Veterinary Medicine, Henan University of Science and Technology, 70 Tianjin Road, Luoyang, Henan 471003, People's Republic of China.
authors: Tao
authors: T
authors: Kong
authors: Xue-Qin
authors: XQ
authors: Hao
authors: Xiao-Bing
authors: XB
authors: Li
authors: Guo-Wen
authors: GW
authors: Liu
authors: Zhi-Gang
authors: ZG
authors: Zhang
authors: Zhi-Jun
authors: ZJ
authors: Yang
authors: Zhe
authors: Z
authors: Wang
authors: Jia
authors: J
authors: Tang
authors: Wei
authors: W
authors: Yang
authors: Jia
authors: J
authors: Sun
mesh_terms: D000085:Acetates / Q000737:chemistry / Q000276:immunology*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000378:metabolism; D000941:Antigens / Q000737:chemistry / Q000276:immunology; D002104:Cadmium / Q000032:analysis / Q000737:chemistry / Q000276:immunology*; D002417:Cattle; D060766:Drinking Water / Q000032:analysis / Q000737:chemistry; D004492:Edetic Acid / Q000031:analogs & derivatives* / Q000737:chemistry / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005029:Ethylenediamines / Q000737:chemistry / Q000276:immunology*; D006825:Hybridomas / Q000166:cytology / Q000378:metabolism; D007114:Immunization; D051379:Mice; D015394:Molecular Structure; D015203:Reproducibility of Results; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000276:immunology*; D013056:Spectrophotometry, Ultraviolet
chemical_list: C509200:1-(4-aminobenzyl)ethylenediamine-N,N,N',N'-tetraacetate; D000085:Acetates; D000911:Antibodies, Monoclonal; D000941:Antigens; D060766:Drinking Water; D005029:Ethylenediamines; D002104:Cadmium; D012710:Serum Albumin, Bovine; C057690:aminobenzyl-EDTA; D004492:Edetic Acid