Development of ELISA for detection of mercury based on specific monoclonal antibodies against mercury-chelate

Biol Trace Elem Res. 2011 Dec;144(1-3):854-64. doi: 10.1007/s12011-011-9031-z. Epub 2011 Mar 25.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal / chemistry*
  • Antibodies, Monoclonal / isolation & purification
  • Antibody Affinity
  • Antibody Specificity
  • Cell Line, Tumor
  • Chelating Agents / chemistry*
  • China
  • Cross Reactions
  • Edetic Acid / analogs & derivatives
  • Edetic Acid / chemistry
  • Environmental Monitoring / methods
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Female
  • Hemocyanins / chemistry
  • Hybridomas
  • Indicators and Reagents
  • Mercury / analysis*
  • Mercury / immunology
  • Metals / analysis
  • Mice
  • Mice, Inbred BALB C
  • Reference Standards
  • Rivers
  • Serum Albumin, Bovine / chemistry
  • Water Pollutants, Chemical / analysis

Substances

  • Antibodies, Monoclonal
  • Chelating Agents
  • Indicators and Reagents
  • Metals
  • Water Pollutants, Chemical
  • Serum Albumin, Bovine
  • aminobenzyl-EDTA
  • Hemocyanins
  • Edetic Acid
  • keyhole-limpet hemocyanin
  • Mercury