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Sensitive detection of atrazine in tap water using TELISA

Qie, Zhiwei; Bai, Jialei; Xie, Bin LU ; Yuan, Lin; Song, Nan; Peng, Yuan; Fan, Xianjun; Zhou, Huanying; Chen, Fengchun and Li, Shuang, et al. (2015) In Analyst 140(15). p.5220-5226
Abstract
A highly sensitive flow injection analysis (FIA)-based thermal enzyme-linked immunoassay, TELISA, was developed for the rapid detection of atrazine (ATZ) in tap water. ATZ and beta-lactamase-labeled ATZ were employed in a competitive immunoassay using a monoclonal antibody (mAb). After the off-column liquid-phase competition, the mAb was captured on the Protein G Sepharose (TM) 4 Fast Flow (PGSFF) column support material. Injected beta-lactamase substrate ampicillin was degraded by the column-bound ATZ-beta-lactamase, generating a detectable heat signal. Several assay parameters were optimized, including substrate concentration, flow rates and regeneration conditions, as well as the mAb and ATZ-beta dilution ratios and concentrations. The... (More)
A highly sensitive flow injection analysis (FIA)-based thermal enzyme-linked immunoassay, TELISA, was developed for the rapid detection of atrazine (ATZ) in tap water. ATZ and beta-lactamase-labeled ATZ were employed in a competitive immunoassay using a monoclonal antibody (mAb). After the off-column liquid-phase competition, the mAb was captured on the Protein G Sepharose (TM) 4 Fast Flow (PGSFF) column support material. Injected beta-lactamase substrate ampicillin was degraded by the column-bound ATZ-beta-lactamase, generating a detectable heat signal. Several assay parameters were optimized, including substrate concentration, flow rates and regeneration conditions, as well as the mAb and ATZ-beta dilution ratios and concentrations. The assay linear range was 0.73-4.83 ng mL(-1) with a detection limit of 0.66 ng mL(-1). An entire heat signal requires 10 min for generation, and the cycle time is less than 40 min. The results were reproducible and stable. ATZ-spiked tap water samples exhibited a recovery rate of 103%-116%, which correlated with the UHPLC-MS/MS measurements. We attributed this significant increase in sensitivity over our previously published work to the following factors: (i) the capture of already-formed immune complexes on the column via immobilized Protein G, which eliminated chemical immobilization of the antibody; (ii) off-column preincubation allows the formation of immune complexes under nearly ideal conditions; and (iii) multiple buffers can be used to, in one case, enhance immune-complex formation and in the other to maximize enzymatic activity. Furthermore, the scheme creates a universal assay platform in which sensing is performed in the off-column incubation and detection after capture in the enzyme thermistor (ET) detector, which opens up the possibility of detecting any antigen for which antibodies were available. (Less)
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publication status
published
subject
in
Analyst
volume
140
issue
15
pages
5220 - 5226
publisher
Royal Society of Chemistry
external identifiers
  • wos:000357810800038
  • scopus:84951748841
ISSN
1364-5528
DOI
10.1039/c5an00636h
language
English
LU publication?
yes
id
47a6b735-2a47-40d0-8405-b34b655ff4b1 (old id 7790945)
date added to LUP
2015-09-03 09:02:36
date last changed
2017-01-22 03:51:00
@article{47a6b735-2a47-40d0-8405-b34b655ff4b1,
  abstract     = {A highly sensitive flow injection analysis (FIA)-based thermal enzyme-linked immunoassay, TELISA, was developed for the rapid detection of atrazine (ATZ) in tap water. ATZ and beta-lactamase-labeled ATZ were employed in a competitive immunoassay using a monoclonal antibody (mAb). After the off-column liquid-phase competition, the mAb was captured on the Protein G Sepharose (TM) 4 Fast Flow (PGSFF) column support material. Injected beta-lactamase substrate ampicillin was degraded by the column-bound ATZ-beta-lactamase, generating a detectable heat signal. Several assay parameters were optimized, including substrate concentration, flow rates and regeneration conditions, as well as the mAb and ATZ-beta dilution ratios and concentrations. The assay linear range was 0.73-4.83 ng mL(-1) with a detection limit of 0.66 ng mL(-1). An entire heat signal requires 10 min for generation, and the cycle time is less than 40 min. The results were reproducible and stable. ATZ-spiked tap water samples exhibited a recovery rate of 103%-116%, which correlated with the UHPLC-MS/MS measurements. We attributed this significant increase in sensitivity over our previously published work to the following factors: (i) the capture of already-formed immune complexes on the column via immobilized Protein G, which eliminated chemical immobilization of the antibody; (ii) off-column preincubation allows the formation of immune complexes under nearly ideal conditions; and (iii) multiple buffers can be used to, in one case, enhance immune-complex formation and in the other to maximize enzymatic activity. Furthermore, the scheme creates a universal assay platform in which sensing is performed in the off-column incubation and detection after capture in the enzyme thermistor (ET) detector, which opens up the possibility of detecting any antigen for which antibodies were available.},
  author       = {Qie, Zhiwei and Bai, Jialei and Xie, Bin and Yuan, Lin and Song, Nan and Peng, Yuan and Fan, Xianjun and Zhou, Huanying and Chen, Fengchun and Li, Shuang and Ning, Baoan and Gao, Zhixian},
  issn         = {1364-5528},
  language     = {eng},
  number       = {15},
  pages        = {5220--5226},
  publisher    = {Royal Society of Chemistry},
  series       = {Analyst},
  title        = {Sensitive detection of atrazine in tap water using TELISA},
  url          = {http://dx.doi.org/10.1039/c5an00636h},
  volume       = {140},
  year         = {2015},
}