title: Development of ELISA for metallothionein-II allows determination of heavy metal pollution of fresh water.
abstract: Metallothionein (MT), a metal-binding protein induced primarily by heavy metals in vertebrates, is considered a biomarker for environmental heavy-metal contamination. To investigate heavy metal pollution in the freshwater environment, MT-I and MT-II were purified from livers of crucian carp (Carassius carassius) by gel exclusion chromatography and ion exchange chromatography. To detect the purified MT-II, a specific monoclonal antibody (mAb) against crucian carp MT-II was produced from the hybridoma strains by cell-cell fusion. By using Enzyme-Linked Immunosorbent Assay (ELISA) with this mAb, the purified crucian carp MT-II was detected with a high specificity and sensitivity. There was a good correlation between the amount of MT-II in carp livers and the concentration of heavy metals in water. ELISA was then used to evaluated the degree of heavy metal pollution in two freshwater systems. The results indicate that the MT-II content in carp liver tissue can be used as an indicator of environmental heavy-metal pollution.
authors.affiliation: National Laboratory of Protein Engineering and Plant Genetic Engineering, College of Life Sciences, Peking University, Beijing 100871, China.
authors.forename: Ming-ming
authors.initials: MM
authors.lastname: Chu
authors.forename: Zhen-quan
authors.initials: ZQ
authors.lastname: Guo
authors.forename: Norio
authors.initials: N
authors.lastname: Muto
authors.forename: Norio
authors.initials: N
authors.lastname: Itoh
authors.forename: Keiichi
authors.initials: K
authors.lastname: Tanaka
authors.forename: Hong-wei
authors.initials: HW
authors.lastname: Ren
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D015415:Biomarkers / Q000032:analysis*; D002347:Carps; D004784:Environmental Monitoring / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D008099:Liver / Q000737:chemistry; D008668:Metallothionein / Q000032:analysis* / Q000378:metabolism; D019216:Metals, Heavy / Q000032:analysis* / Q000378:metabolism; D013054:Spectrophotometry, Atomic; D014873:Water Pollutants / Q000032:analysis* / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D015415:Biomarkers; D019216:Metals, Heavy; D014873:Water Pollutants; D008668:Metallothionein