Abstract
A competitive enzyme-linked immunosorbent assay (ELISA) was developed for the quantitative detection of the organophosphorus insecticide bromophos-ethyl. Four bromophos-ethyl derivatives (haptens) were synthesized and were coupled to carrier proteins through the pesticide thiophosphate group to use as an immunogen or as a coating antigen. Rabbits were immunized with either one of two haptens coupled to bovine serum albumin for production of polyclonal antibodies, and the sera were screened against one of the haptens coupled to ovalbumin. Using the serum with highest titer, an antigen-coated ELISA was developed, which showed an IC50 of 3.9 ng/mL with a detection limit of 0.3 ng/mL (20% inhibition). An antibody-coated ELISA using an enzyme tracer was also developed, which showed an IC50 of 6.5 ng/mL with a detection limit of 1.0 ng/mL (20% inhibition). The antibodies showed negligible cross-reactivity with other organophosphorus pesticides except with the insecticides bromophos-methyl and chlorpyrifos in the antibody-coated assay only. Recoveries of bromophosethyl from fortified crop and water samples ranged from 82 to 128% and from 95 to 127%, respectively.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 6675-6682 |
| Number of pages | 8 |
| Journal | Journal of Agricultural and Food Chemistry |
| Volume | 50 |
| Issue number | 23 |
| DOIs | |
| State | Published - Nov 6 2002 |
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Keywords
- Bromophos-ethyl
- ELISA
- Immunoassay
- Insecticide
ASJC Scopus subject areas
- Agricultural and Biological Sciences (miscellaneous)
- Food Science
- Chemistry (miscellaneous)
Cite this
Development of an enzyme-linked immunosorbent assay for the organophosphorus insecticide bromophos-ethyl. / Kim, Kwang Ok; Kim, Yoo Jung; Lee, Yong Tae; Hammock, Bruce D.; Lee, Hye Sung.
In: Journal of Agricultural and Food Chemistry, Vol. 50, No. 23, 06.11.2002, p. 6675-6682.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Development of an enzyme-linked immunosorbent assay for the organophosphorus insecticide bromophos-ethyl
AU - Kim, Kwang Ok
AU - Kim, Yoo Jung
AU - Lee, Yong Tae
AU - Hammock, Bruce D.
AU - Lee, Hye Sung
PY - 2002/11/6
Y1 - 2002/11/6
N2 - A competitive enzyme-linked immunosorbent assay (ELISA) was developed for the quantitative detection of the organophosphorus insecticide bromophos-ethyl. Four bromophos-ethyl derivatives (haptens) were synthesized and were coupled to carrier proteins through the pesticide thiophosphate group to use as an immunogen or as a coating antigen. Rabbits were immunized with either one of two haptens coupled to bovine serum albumin for production of polyclonal antibodies, and the sera were screened against one of the haptens coupled to ovalbumin. Using the serum with highest titer, an antigen-coated ELISA was developed, which showed an IC50 of 3.9 ng/mL with a detection limit of 0.3 ng/mL (20% inhibition). An antibody-coated ELISA using an enzyme tracer was also developed, which showed an IC50 of 6.5 ng/mL with a detection limit of 1.0 ng/mL (20% inhibition). The antibodies showed negligible cross-reactivity with other organophosphorus pesticides except with the insecticides bromophos-methyl and chlorpyrifos in the antibody-coated assay only. Recoveries of bromophosethyl from fortified crop and water samples ranged from 82 to 128% and from 95 to 127%, respectively.
AB - A competitive enzyme-linked immunosorbent assay (ELISA) was developed for the quantitative detection of the organophosphorus insecticide bromophos-ethyl. Four bromophos-ethyl derivatives (haptens) were synthesized and were coupled to carrier proteins through the pesticide thiophosphate group to use as an immunogen or as a coating antigen. Rabbits were immunized with either one of two haptens coupled to bovine serum albumin for production of polyclonal antibodies, and the sera were screened against one of the haptens coupled to ovalbumin. Using the serum with highest titer, an antigen-coated ELISA was developed, which showed an IC50 of 3.9 ng/mL with a detection limit of 0.3 ng/mL (20% inhibition). An antibody-coated ELISA using an enzyme tracer was also developed, which showed an IC50 of 6.5 ng/mL with a detection limit of 1.0 ng/mL (20% inhibition). The antibodies showed negligible cross-reactivity with other organophosphorus pesticides except with the insecticides bromophos-methyl and chlorpyrifos in the antibody-coated assay only. Recoveries of bromophosethyl from fortified crop and water samples ranged from 82 to 128% and from 95 to 127%, respectively.
KW - Bromophos-ethyl
KW - ELISA
KW - Immunoassay
KW - Insecticide
UR - http://www.scopus.com/inward/record.url?scp=0037032287&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0037032287&partnerID=8YFLogxK
U2 - 10.1021/jf025703+
DO - 10.1021/jf025703+
M3 - Article
C2 - 12405760
AN - SCOPUS:0037032287
VL - 50
SP - 6675
EP - 6682
JO - Journal of Agricultural and Food Chemistry
JF - Journal of Agricultural and Food Chemistry
SN - 0021-8561
IS - 23
ER -