Objective To improve and apply the competitive ELISA method for measuring human urinary metallothionein (UMT).
Methods Human metallothionein(MT)was isolated from an adult liver by SephadexG-75 and Sephadex G-50 chromatographic column, and then used to immunize Balb/c mice for preparing monoclonal antibody(McAb). The competitive ELISA was established with the McAb as the primary antibody. UMT was determined in 298 urine samples from university students.
Results The detection limit of the ELISA was 9 ng/mL. The coefficients of variation for the within-running and between-running were 1.78%-6.72% and 5.45%-13.26%, respectively, while the recovery was 99.69%-107.25%. Before physical exercise, UMT concentrations of subjects were(10.58& #177;5.62)ng/μmol Cr, while their UMT concentrations increased significantly after exercising, reached to(19.73& #177;8.65)ng/μmol Cr. UMT levels in smokers(86.58& #177;30.96)ng/μmol Cr were significantly higher than those in non-smokers(14.73& #177;6.57)ng/μmol Cr.
Conclusion The competitive ELISA established was accurate and precise for measuring human UMT. The physical exercising and smoking influenced the UMT levels.
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