Abstract
Epidemiological and toxicological studies often require the analysis of large numbers of samples for biological markers of exposure. The goal of this work was to develop a class-selective ELISA to detect groups of structurally closely related mercapturic acids with small nonpolar S-substituents. An assay was developed with strong recognition for mercapturates including S-benzylmercapturic acid (IC50 = 0.018 μmol/L), S-n-hexylmercapturic acid (IC50 = 0.021 μmol/L), S-phenylmercapturic acid (IC50 = 0.024 μmol/L), and S-cyclohexylmethylmercapturic acid (IC50 = 0.042 μmol/L). The same assay also showed weaker recognition for S-(1-hydroxynaphthal-2-yl)mercapturic acid and S-allylmercapturic acid (IC50 = 1.1 and 1.7 μmol/L, respectively). Subtle modifications to the hapten linker structure of the coating antigen proved to have a strong impact on the selectivity and the specificity of the assay. A slightly modified assay showed high recognition for S-benzylmercapturic acid (IC50 = 0.018 μmol/L) and weaker recognition for seven other mercapturic acids (IC50 = 0.021-10 μmol/L). Strong positive assay responses were detected in 12 urine samples obtained from persons with no known occupational exposure to exogenous electrophilic xenobiotics. Solid phase extraction and cross-reactivity indicated that the presumptive immunoreactive materials were similar in size and polarity to S-benzylmercapturic acid. The assay was more selective to mercapturic acids than the spectrophotometric thioether assay.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 5913-5923 |
| Number of pages | 11 |
| Journal | Journal of Agricultural and Food Chemistry |
| Volume | 48 |
| Issue number | 12 |
| DOIs | |
| State | Published - 2000 |
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Keywords
- Biological monitoring
- ELISA
- Mercapturic acid
- Urine
ASJC Scopus subject areas
- Agricultural and Biological Sciences (miscellaneous)
- Food Science
- Chemistry (miscellaneous)
Cite this
Development of a class-selective enzyme-linked immunosorbent assay for mercapturic acids in human urine. / Lohse, Christian; Jaeger, Lynn L.; Staimer, Norbert; Sanborn, Jim R.; Daniel Jones, A.; Langó, József; Gee, Shirley J.; Hammock, Bruce D.
In: Journal of Agricultural and Food Chemistry, Vol. 48, No. 12, 2000, p. 5913-5923.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Development of a class-selective enzyme-linked immunosorbent assay for mercapturic acids in human urine
AU - Lohse, Christian
AU - Jaeger, Lynn L.
AU - Staimer, Norbert
AU - Sanborn, Jim R.
AU - Daniel Jones, A.
AU - Langó, József
AU - Gee, Shirley J.
AU - Hammock, Bruce D.
PY - 2000
Y1 - 2000
N2 - Epidemiological and toxicological studies often require the analysis of large numbers of samples for biological markers of exposure. The goal of this work was to develop a class-selective ELISA to detect groups of structurally closely related mercapturic acids with small nonpolar S-substituents. An assay was developed with strong recognition for mercapturates including S-benzylmercapturic acid (IC50 = 0.018 μmol/L), S-n-hexylmercapturic acid (IC50 = 0.021 μmol/L), S-phenylmercapturic acid (IC50 = 0.024 μmol/L), and S-cyclohexylmethylmercapturic acid (IC50 = 0.042 μmol/L). The same assay also showed weaker recognition for S-(1-hydroxynaphthal-2-yl)mercapturic acid and S-allylmercapturic acid (IC50 = 1.1 and 1.7 μmol/L, respectively). Subtle modifications to the hapten linker structure of the coating antigen proved to have a strong impact on the selectivity and the specificity of the assay. A slightly modified assay showed high recognition for S-benzylmercapturic acid (IC50 = 0.018 μmol/L) and weaker recognition for seven other mercapturic acids (IC50 = 0.021-10 μmol/L). Strong positive assay responses were detected in 12 urine samples obtained from persons with no known occupational exposure to exogenous electrophilic xenobiotics. Solid phase extraction and cross-reactivity indicated that the presumptive immunoreactive materials were similar in size and polarity to S-benzylmercapturic acid. The assay was more selective to mercapturic acids than the spectrophotometric thioether assay.
AB - Epidemiological and toxicological studies often require the analysis of large numbers of samples for biological markers of exposure. The goal of this work was to develop a class-selective ELISA to detect groups of structurally closely related mercapturic acids with small nonpolar S-substituents. An assay was developed with strong recognition for mercapturates including S-benzylmercapturic acid (IC50 = 0.018 μmol/L), S-n-hexylmercapturic acid (IC50 = 0.021 μmol/L), S-phenylmercapturic acid (IC50 = 0.024 μmol/L), and S-cyclohexylmethylmercapturic acid (IC50 = 0.042 μmol/L). The same assay also showed weaker recognition for S-(1-hydroxynaphthal-2-yl)mercapturic acid and S-allylmercapturic acid (IC50 = 1.1 and 1.7 μmol/L, respectively). Subtle modifications to the hapten linker structure of the coating antigen proved to have a strong impact on the selectivity and the specificity of the assay. A slightly modified assay showed high recognition for S-benzylmercapturic acid (IC50 = 0.018 μmol/L) and weaker recognition for seven other mercapturic acids (IC50 = 0.021-10 μmol/L). Strong positive assay responses were detected in 12 urine samples obtained from persons with no known occupational exposure to exogenous electrophilic xenobiotics. Solid phase extraction and cross-reactivity indicated that the presumptive immunoreactive materials were similar in size and polarity to S-benzylmercapturic acid. The assay was more selective to mercapturic acids than the spectrophotometric thioether assay.
KW - Biological monitoring
KW - ELISA
KW - Mercapturic acid
KW - Urine
UR - http://www.scopus.com/inward/record.url?scp=0034524145&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0034524145&partnerID=8YFLogxK
U2 - 10.1021/jf000621b
DO - 10.1021/jf000621b
M3 - Article
C2 - 11312767
AN - SCOPUS:0034524145
VL - 48
SP - 5913
EP - 5923
JO - Journal of Agricultural and Food Chemistry
JF - Journal of Agricultural and Food Chemistry
SN - 0021-8561
IS - 12
ER -