title: Preparation and characterization of monoclonal antibody specific for copper-chelate complex.
abstract: Monoclonal antibody (MAb) against chelated Cu(2+) was developed. The conjugate employed as immunogens in BALB/c mice to raise antibodies was synthesized using Cu(2+) coupling to keyhole limpet hemocyanin (KLH) via a bifunctional chelator (2-(4-isothiocyanatobenzyl)-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid, p-SCN-Bn-DOTA). One of the hybridomas secreting antibody that bound tightly to Cu(2+)-ethylenediamine tetraacetic acid (EDTA) complex but not to metal-free EDTA was isolated from the fusion between murine splenocytes and SP2/0 myeloma cells. The performance of a competitive enzyme-linked immunosorbent assay (ELISA) was assessed for its sensitivity to changes in pH and ionic strength. The cross-reactivities (CR) with other metals were below 1%, except for Hg(2+) with a CR of 7.19 %. The IC(50) value achieved for copper was 0.89 μg/mL, showing a detection range of 0.25-8.13 μg/mL and the lowest detection limit of 0.032 μg/mL. The concentration of Cu(2+) in environmental water and serum samples obtained by ELISA correlated well with Atomic Absorption Spectrometry (AAS), and the mean recovery was 93.7%. These results indicated that ELISA could be a convenient analytical tool for monitoring copper in drinking water and human serum.
authors.affiliation: Key Lab of Monitoring and Management of Crop Diseases and Pest Insects, Ministry of Agriculture, College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China.
authors.forename: Fengquan
authors.initials: F
authors.lastname: Liu
authors.forename: Yang
authors.initials: Y
authors.lastname: Lou
authors.forename: Xiuying
authors.initials: X
authors.lastname: Shi
authors.forename: Huiming
authors.initials: H
authors.lastname: Wang
authors.forename: Xiaoxia
authors.initials: X
authors.lastname: Zhu
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D002614:Chelating Agents / Q000737:chemistry*; D003300:Copper / Q000097:blood / Q000737:chemistry* / Q000276:immunology*; D003429:Cross Reactions / Q000276:immunology; D060766:Drinking Water / Q000032:analysis; D004492:Edetic Acid / Q000031:analogs & derivatives / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006433:Hemocyanins / Q000737:chemistry; D006573:Heterocyclic Compounds, 1-Ring / Q000737:chemistry; D006825:Hybridomas; D006863:Hydrogen-Ion Concentration; D017879:Isothiocyanates / Q000737:chemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D009994:Osmolar Concentration; D015203:Reproducibility of Results; D044967:Serum / Q000737:chemistry / Q000276:immunology; D013054:Spectrophotometry, Atomic
chemical_list: D000911:Antibodies, Monoclonal; D002614:Chelating Agents; D060766:Drinking Water; D006573:Heterocyclic Compounds, 1-Ring; D017879:Isothiocyanates; C119668:isothiocyanobenzyl ethylenediamine tetracetic acid; C071349:1,4,7,10-tetraazacyclododecane- 1,4,7,10-tetraacetic acid; D003300:Copper; D006433:Hemocyanins; D004492:Edetic Acid; C032808:keyhole-limpet hemocyanin