Development of enzyme-linked immunosorbent assay (ELISA) for glutathione S-transferase (GST-S) protein in the intertidal copepod Tigriopus japonicus and its application for environmental monitoring

Chemosphere. 2013 Nov;93(10):2458-66. doi: 10.1016/j.chemosphere.2013.08.077. Epub 2013 Oct 7.


To utilize the GST-S protein as a useful biomarker for environmental contamination, we developed a polyclonal antibody-based enzyme-linked immunosorbent assay (ELISA) in the intertidal copepod Tigriopus japonicus. Two polyclonal antibodies, TJ-GST-S1 and TJ-GST-S2, were raised against two TJ-GST-S synthetic peptides. Also a recombinant TJ-GST-S protein was purified as a standard for ELISA development. Each polyclonal antibody was tested by Western blot analysis and indirect ELISA. Of two polyclonal antibodies, TJ-GST-S2 ELISA was further employed due to its wide range of detection and the limit of specificity compared to those of TJ-GST-S1 ELISA system. After exposure to 4 metals (Ag, As, Cd, and Cu) to T. japonicus, the amount of TJ-GST-S protein was significantly elevated in a concentration-dependent manner. Also, TJ-GST-S protein was upregulated at relative high concentrations of B[α]P, PCB, and TBT. In this paper, we suggest that T. japonicas ELISA for TJ-GST-S2 is useful as a potential indicator system for marine contaminants.

Keywords: Contaminant; Copepod; Enzyme-linked immunosorbent assay; Glutathione S-transferase; Tigriopus japonicus.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Copepoda / metabolism*
  • Environmental Monitoring / methods*
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Gene Expression
  • Glutathione Transferase / genetics
  • Glutathione Transferase / metabolism*
  • Water Pollutants, Chemical / toxicity*


  • Water Pollutants, Chemical
  • Glutathione Transferase