Instrumental Aspects of Food Analysis by Electrochemical Methods

Coltro, Wendell K. T.; Mora, María F.; Garcı́a, Carlos D.

doi:10.1002/9781118684030.ch16

Cited by 1 publication

(1 citation statement)

References 187 publications

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“…The most frequently used instrumental detection methods for DA are liquid chromatography-tandem mass spectrometry (LC-MS/MS), liquid chromatography ultraviolet detection (LC-UV), biosensors, and immunoassays [ 3 , 10 , 11 ]. Instrument-based methods are sensitive and they can be used simultaneously for the analysis of multiples toxins, however, the drawbacks include equipment complexity, exhibit incompatibility, costly and prolonged when use for the real samples [ 12 ]. ELISA is an exceedingly sensitive immunoassay using enzyme-conjugated antibodies, with antigen or antibodies captured to a solid binding support such as ELISA plate [ 6 , 13 , 14 , 15 ].…”

Section: Introductionmentioning

confidence: 99%

The Preparation and Identification of a Monoclonal Antibody against Domoic Acid and Establishment of Detection by Indirect Competitive ELISA

Saeed

Ling

Yuan

et al. 2017

Toxins

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Domoic acid (DA) is a potent toxin, marine biotoxin, and primarily produced by Pseudo-nitzschia. The DA hapten was coupled with bovine serum albumin (BSA), and ovalbumin (OVA) as carrier proteins. DA-BSA conjugate was used as immunogen and DA-OVA as coating antigen. Cell fusion between spleen cells and sp2/0 myeloma cells developed 1C3 hybridoma clone producing 1C3 monoclonal antibody (mAb). Hybridoma was injected into the mice to produce ascites, and further purified by caprylic acid/ammonium sulfate method. The mAb was of IgG3 subclass, and was specific to DA with high affinity (2.5 × 108 L/mol). Moreover, western blot exhibited significant specificity to the DA antigens. Indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) showed DA working range of 0.006–0.2 ng/mL. The IC50 was 0.03 ng/mL with low limit of detection (LOD) of 0.006 ng/mL. Average DA recovery from spiked shellfish extract was 100.56 ± 2.8% with the coefficient variation of 0.01–0.1%. Hence, mAb producing 1C3 hybridoma was successfully developed and could be used to detect DA in contaminated samples.

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