Polyclonal antibodies to domoic acid, and their use in immunoassays for domoic acid in sea water and shellfish

Garthwaite, Ian; Ross, Kathryn; Miles, Christopher O.; Hansen, Richard P.; Foster, David L.; Wilkins, Alistair L.; Towers, N. R.

doi:10.1002/(sici)1522-7189(199805/08)6:3/4<93::aid-nt15>3.0.co;2-9

Cited by 80 publications

(42 citation statements)

References 13 publications

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“…Standards serially diluted in Nanopure water and samples were loaded in duplicate or triplicate into wells of a 96-well plate coated with BSA with carboxyl-linked DA antigen (18). Only Nanopure water was added to wells for determination of A max .…”

Section: Methodsmentioning

confidence: 99%

“…Only Nanopure water was added to wells for determination of A max . Fifty microliters of anti-DA antibody [generated by injecting a rabbit with an imine-linked DA antigen (18)] was added to each well except blank wells, to which 50 μL of blocking buffer (3% nonfat dry milk in PBS solution) was added. The plate was sealed with plastic sealer, covered with foil, and gently mixed on an orbital shaker for 1 h at room temperature.…”

Section: Methodsmentioning

confidence: 99%

“…Pseudonitzschia turgidula and the smaller P. cf granii were present in low concentrations (<1,000 cells L No other Pseudonitzschia species were observed during this study. We collected 1-L water samples at each depth and measured particulate DA (pDA) on fresh samples on board the ship using an indirect competitive ELISA (cELISA) method (18). Particulate DA was measured at 5 m depth over 6 consecutive days (Fig.…”

Section: Measurement Of Cellular Domoic Acid In Subarctic Pacific Surmentioning

confidence: 99%

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Iron enrichment stimulates toxic diatom production in high-nitrate, low-chlorophyll areas

Trick

Bill

Cochlan

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

107

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Oceanic high-nitrate, low-chlorophyll environments have been highlighted for potential large-scale iron fertilizations to help mitigate global climate change. Controversy surrounds these initiatives, both in the degree of carbon removal and magnitude of ecosystem impacts. Previous open ocean enrichment experiments have shown that iron additions stimulate growth of the toxigenic diatom genus Pseudonitzschia . Most Pseudonitzschia species in coastal waters produce the neurotoxin domoic acid (DA), with their blooms causing detrimental marine ecosystem impacts, but oceanic Pseudonitzschia species are considered nontoxic. Here we demonstrate that the sparse oceanic Pseudonitzschia community at the high-nitrate, low-chlorophyll Ocean Station PAPA (50° N, 145° W) produces approximately 200 pg DA L −1 in response to iron addition, that DA alters phytoplankton community structure to benefit Pseudonitzschia , and that oceanic cell isolates are toxic. Given the negative effects of DA in coastal food webs, these findings raise serious concern over the net benefit and sustainability of large-scale iron fertilizations.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Measurement Of Cellular Domoic Acid In Subarctic Pacific Surmentioning

confidence: 99%

See 1 more Smart Citation

Iron enrichment stimulates toxic diatom production in high-nitrate, low-chlorophyll areas

Trick

Bill

Cochlan

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

107

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“…The current work focuses on the development of a disposable electrochemical immunosensor for the detection of DA in mussel tissue using a polyclonal antibody produced by Toxicology and Food Safety Research (New Zealand) (Garthwaite et al, 1998). Spectrophotometric ELISA was primarily used as a development tool, prior to transfer of the DA assay onto screen-printed electrodes (SPEs).…”

Section: Introductionmentioning

confidence: 99%

Disposable immunosensor for the determination of domoic acid in shellfish

Micheli

Radoi

Guarrina

et al. 2004

Biosensors and Bioelectronics

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The construction of an electrochemical immunosensor coupled to differential pulse voltammetry (DPV) for the detection of domoic acid (DA), a neurotoxic aminoacid responsible for the human syndrome known as "Amnesic Shellfish Poisoning" (ASP), is proposed here.The method involves the use of disposable screen-printed electrodes (SPEs) for the immunosensor development based on a "competitive indirect test". Domoic acid conjugated to bovine serum albumin (BSA-DA) was coated onto the working electrode of the SPE, followed by incubation with sample (or standard toxin) and anti-DA antibody. An anti-goat IgG-alkaline phosphatase (AP) conjugate was used for signal generation. A spectrophotometric enzyme-linked immunosorbent assay (ELISA) was used in a preliminary phase of development, prior to transferring the assay to the SPEs.Results showed a detection limit equal to 5 ng/ml of toxin. The electrochemical system is simple and cost-effective due to the disposable nature of the SPEs, and the analysis time is 150 min, shorter than that for the spectrophotometric method.The suitability of the assay for DA quantification in mussels was also evaluated. Samples were spiked with DA before and after the sample treatment to study the extraction efficiency and the matrix effect, respectively. After treatment, samples were analysed using a 1:250 v/v dilution in PBS-M (phosphate saline buffer pH 7.4 + CH 3 OH 10%) to minimise the matrix effect and allow for the detection of 20 g/g of DA in mussel tissue. This represents the maximum acceptable limit defined by the Food and Drug Administration [Compliance Programme 7303.842. Guidance Levels, Table 3, p. 248, http://www.fda.org]. The optimised ELISA systems were then used, in parallel with a conventional HPLC method, to detect and confirm DA in shellfish extract in order to verify the performance of the electrochemical system. Very good recoveries were obtained, demonstrating the suitability of the proposed assay for accurate determination of the DA concentration in mussel samples.

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“…As antibodies produced against DA have led to the production of enzyme linked immunoabsorbent assays for DA, use of certain antibodies have led to the synthesis of another detection method, direct competitive enzyme-linked immunosorbent assay (cELISA) (Smith et al, 1994;Garthwaite et al, 1998;Kleivdal et al, 2007). The assay has now been commercialized and serves mainly for the detection of DA in shellfish (Kleivdal et al, 2007).…”

Section: Monitoring Damentioning

confidence: 99%

The Photooxidation of Domoic Acid

Parekh¹

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Polyclonal antibodies to domoic acid, and their use in immunoassays for domoic acid in sea water and shellfish

Cited by 80 publications

References 13 publications

Iron enrichment stimulates toxic diatom production in high-nitrate, low-chlorophyll areas

Iron enrichment stimulates toxic diatom production in high-nitrate, low-chlorophyll areas

Disposable immunosensor for the determination of domoic acid in shellfish

The Photooxidation of Domoic Acid

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