TY - JOUR
T1 - Quantitative microarray pesticide analysis
AU - Belleville, Erik
AU - Dufva, Martin
AU - Aamand, Jens
AU - Bruun, Leif
AU - Clausen, Liselotte
AU - Christensen, Claus B.V.
N1 - Funding Information:
This work was financially supported by the Danish Research Council (SUE project no. 2027-00-0005 and DABIC project no. 2014-00-0003).
PY - 2004/3
Y1 - 2004/3
N2 - To replace a pesticide immunoassay based on microtiter plates, we have developed a quantitative, competitive microarray immunoassay, which permits rapid and highly sensitive quantification of the dichlobenil degradation product 2,6-dichlorobenzamide (BAM), and the prominently used herbicide atrazine. The pesticide analysis is based on the competitive binding of fluorescence conjugated monoclonal antibodies (mAb) to their respective analytes. Lowest detection limits were calculated to 1 ng/l (5 pM) for BAM and 3 ng/l (10 pM) for atrazine. Corresponding IC
50 values were, 10 ng/l (50 pM) for BAM and 34 ng/l (160 pM) for atrazin, respectively. In comparison to the existing microtiter plate immunoassay, the microarray was found to be up to 20-fold more sensitive. Compared to the gas chromatography with mass spectroscopy (GCMS) analysis performed on more than 1000-fold concentrated samples, the microarray-based immunoassay was even 10-fold more sensitive using non-concentrated samples. Measuring both analytes simultaneously did not affect assay sensitivity compared to single analyte quantification. Besides a gain in sensitivity and the possibility of multiplex quantification, assay times and assay complexity were reduced drastically with the microarray platform compared to the microtiter plate immunoassay and GCMS, suggesting that the microarray based immunoassay is a viable method for measuring picomolar amounts of analytes, e.g. clinically relevant analytes.
AB - To replace a pesticide immunoassay based on microtiter plates, we have developed a quantitative, competitive microarray immunoassay, which permits rapid and highly sensitive quantification of the dichlobenil degradation product 2,6-dichlorobenzamide (BAM), and the prominently used herbicide atrazine. The pesticide analysis is based on the competitive binding of fluorescence conjugated monoclonal antibodies (mAb) to their respective analytes. Lowest detection limits were calculated to 1 ng/l (5 pM) for BAM and 3 ng/l (10 pM) for atrazine. Corresponding IC
50 values were, 10 ng/l (50 pM) for BAM and 34 ng/l (160 pM) for atrazin, respectively. In comparison to the existing microtiter plate immunoassay, the microarray was found to be up to 20-fold more sensitive. Compared to the gas chromatography with mass spectroscopy (GCMS) analysis performed on more than 1000-fold concentrated samples, the microarray-based immunoassay was even 10-fold more sensitive using non-concentrated samples. Measuring both analytes simultaneously did not affect assay sensitivity compared to single analyte quantification. Besides a gain in sensitivity and the possibility of multiplex quantification, assay times and assay complexity were reduced drastically with the microarray platform compared to the microtiter plate immunoassay and GCMS, suggesting that the microarray based immunoassay is a viable method for measuring picomolar amounts of analytes, e.g. clinically relevant analytes.
KW - 2,6-dichlorobenzamide
KW - 2,6-Dichlorobenzamide (BAM)
KW - Anthraquinone
KW - AQ
KW - Atrazine
KW - BAM
KW - Bovine serum albumin
KW - BSA
KW - ELISA
KW - Enzyme-linked immunosorbent assay
KW - GCMS
KW - Immunoassay
KW - Microarray
KW - Monoclonal antibodies
KW - Multiplex quantification
UR - http://www.scopus.com/inward/record.url?scp=1842850827&partnerID=8YFLogxK
U2 - 10.1016/j.jim.2004.01.004
DO - 10.1016/j.jim.2004.01.004
M3 - Article
VL - 286
SP - 219
EP - 229
JO - Journal of Immunological Methods
JF - Journal of Immunological Methods
SN - 0022-1759
IS - 1-2
ER -