Application of a cytochrome P-450 IA1-ELISA in environmental monitoring and toxicological testing of fish.
1. An enzyme-linked immunosorbent assay (ELISA) based on polyclonal rabbit anti-cod cytochrome P-450 IA1 IgG has been developed in our laboratory. 2. The antibodies employed in the ELISA demonstrate good cross-reactivity with the homologous protein in a number of other fish species, giving cross-reacting protein bands o 54-59 kDa in liver microsomes, as determined by Western blotting. 3. The ELISA technique has been used in numerous experiments with both field collected and laboratory exposed fish of different species, showing good correlation with contaminant exposure. 4. In some instances of PCB exposure where the classical P-450 IA1 monooxygenase assay 7-ethoxyresorufin O-deethylase (EROD) failed to reveal any induction, the ELISA technique demonstrated increased levels of P-450 IA1 protein, indicating inhibiting effects of the PCBs on EROD measurement. 5. In tissues like gill filaments, and in whole cod larvae, where EROD activity is barely detectable, if at all, the ELISA technique showed induction after exposure to a water soluble fraction (WSF) of North Sea crude oil. 6. The results reviewed indicate the usefulness of the ELISA technique to allow rapid screening of a large number of samples, and especially when catalytic measurement is difficult or impossible due to (a) small sample or tissue size, (b) loss of activities in bad storage conditions, or (c) presence of compounds inhibiting activity.
Department of Biochemistry, University of Bergen, Norway.
A
A
Goksøyr
H E
HE
Larsen
A M
AM
Husøy
D000818:Animals; D003577:Cytochrome P-450 Enzyme System / Q000187:drug effects* / Q000276:immunology; D004784:Environmental Monitoring / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay; D005399:Fishes / Q000378:metabolism*; D007074:Immunoglobulin G / Q000032:analysis; D014874:Water Pollutants, Chemical / Q000633:toxicity*
D007074:Immunoglobulin G; D014874:Water Pollutants, Chemical; D003577:Cytochrome P-450 Enzyme System