Optimization of a charge coupled device imaging enzyme linked immuno sorbent assay and supports for the simultaneous determination of multiple 2,4-D samples
(1997) In Analytica Chimica Acta 347(1-2). p.87-93- Abstract
A chemiluminescent microformat enzyme linked immune sorbent assay (ELISA) has been optimized for the simultaneous determination of multiple 2,4-dichlorophenoxyacetic acid (2,4-D) samples. The competitive immunoassay employed a 2,4-D-BSA conjugate, anti-2,4-D monoclonal antibodies and alkaline phosphatase (AP) labelled anti-mouse IgG. The bound AP conjugate was determined by quantitating the chemiluminescence emission from the enzymatic decomposition of the luminogenic substrate, CSPD, by AP using a cooled charge coupled device (CCD) camera. The detection limit for the simultaneous determination of multiple samples was 4.3x10-10 M corresponding to 96 pg ml-1 or 192 fg well with a coefficient of variation (CV, %) of... (More)
A chemiluminescent microformat enzyme linked immune sorbent assay (ELISA) has been optimized for the simultaneous determination of multiple 2,4-dichlorophenoxyacetic acid (2,4-D) samples. The competitive immunoassay employed a 2,4-D-BSA conjugate, anti-2,4-D monoclonal antibodies and alkaline phosphatase (AP) labelled anti-mouse IgG. The bound AP conjugate was determined by quantitating the chemiluminescence emission from the enzymatic decomposition of the luminogenic substrate, CSPD, by AP using a cooled charge coupled device (CCD) camera. The detection limit for the simultaneous determination of multiple samples was 4.3x10-10 M corresponding to 96 pg ml-1 or 192 fg well with a coefficient of variation (CV, %) of 12.5%. The linear range of the assay was 4.5 x 10-7-4.5 x 10-10 M. The ability of gold coated silicon wafers and glass capillaries to serve as solid phase supports in the imaging ELISA was investigated. The highly reflective gold surfaces improved both the linear range and the sensitivity of the assay, as compared to thick-film patterned surfaces. The capillary supports, on the other hand, lead to a reduction in the linear range and the sensitivity of the assay, as compared to the thick-film patterned surfaces. Initial studies indicate that the capillaries guide the light and may provide a built-in mechanism for collecting the emitted light. Strategies for further development of support materials for imaging-based detectors will be discussed.
(Less)
- author
- Dzgoev, A. LU ; Mecklenburg, M. ; Xie, B. LU ; Miyabayashi, A. ; Larsson, P. O. LU and Danielsson, B. LU
- organization
- publishing date
- 1997-07-30
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- 2,4-D, Array, Charge coupled devic-camera, CSPD, Enzyme linked immuno sorbent assay, Herbicide, Imaging, Multisample, Support, Thick-film
- in
- Analytica Chimica Acta
- volume
- 347
- issue
- 1-2
- pages
- 7 pages
- publisher
- Elsevier
- external identifiers
-
- scopus:0030879213
- ISSN
- 0003-2670
- DOI
- 10.1016/S0003-2670(97)00002-0
- language
- English
- LU publication?
- yes
- id
- 4a8de895-2c04-426e-b91c-52d0b771cf42
- date added to LUP
- 2021-10-27 13:09:14
- date last changed
- 2022-02-02 00:55:11
@article{4a8de895-2c04-426e-b91c-52d0b771cf42,
abstract = {{<p>A chemiluminescent microformat enzyme linked immune sorbent assay (ELISA) has been optimized for the simultaneous determination of multiple 2,4-dichlorophenoxyacetic acid (2,4-D) samples. The competitive immunoassay employed a 2,4-D-BSA conjugate, anti-2,4-D monoclonal antibodies and alkaline phosphatase (AP) labelled anti-mouse IgG. The bound AP conjugate was determined by quantitating the chemiluminescence emission from the enzymatic decomposition of the luminogenic substrate, CSPD, by AP using a cooled charge coupled device (CCD) camera. The detection limit for the simultaneous determination of multiple samples was 4.3x10<sup>-10</sup> M corresponding to 96 pg ml<sup>-1</sup> or 192 fg well with a coefficient of variation (CV, %) of 12.5%. The linear range of the assay was 4.5 x 10<sup>-7</sup>-4.5 x 10<sup>-10</sup> M. The ability of gold coated silicon wafers and glass capillaries to serve as solid phase supports in the imaging ELISA was investigated. The highly reflective gold surfaces improved both the linear range and the sensitivity of the assay, as compared to thick-film patterned surfaces. The capillary supports, on the other hand, lead to a reduction in the linear range and the sensitivity of the assay, as compared to the thick-film patterned surfaces. Initial studies indicate that the capillaries guide the light and may provide a built-in mechanism for collecting the emitted light. Strategies for further development of support materials for imaging-based detectors will be discussed.</p>}},
author = {{Dzgoev, A. and Mecklenburg, M. and Xie, B. and Miyabayashi, A. and Larsson, P. O. and Danielsson, B.}},
issn = {{0003-2670}},
keywords = {{2,4-D; Array; Charge coupled devic-camera; CSPD; Enzyme linked immuno sorbent assay; Herbicide; Imaging; Multisample; Support; Thick-film}},
language = {{eng}},
month = {{07}},
number = {{1-2}},
pages = {{87--93}},
publisher = {{Elsevier}},
series = {{Analytica Chimica Acta}},
title = {{Optimization of a charge coupled device imaging enzyme linked immuno sorbent assay and supports for the simultaneous determination of multiple 2,4-D samples}},
url = {{http://dx.doi.org/10.1016/S0003-2670(97)00002-0}},
doi = {{10.1016/S0003-2670(97)00002-0}},
volume = {{347}},
year = {{1997}},
}