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1.
Talanta ; 156-157: 55-63, 2016 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-27260435

RESUMO

Research in biosensing approaches as alternative techniques for food diagnostics for the detection of chemical contaminants and foodborne pathogens has increased over the last twenty years. The key component of such tests is the biorecognition element whereby polyclonal or monoclonal antibodies still dominate the market. Traditionally the screening of sera or cell culture media for the selection of polyclonal or monoclonal candidate antibodies respectively has been performed by enzyme immunoassays. For niche toxin compounds, enzyme immunoassays can be expensive and/or prohibitive methodologies for antibody production due to limitations in toxin supply for conjugate production. Automated, self-regenerating, chip-based biosensors proven in food diagnostics may be utilised as rapid screening tools for antibody candidate selection. This work describes the use of both single channel and multi-channel surface plasmon resonance (SPR) biosensors for the selection and characterisation of antibodies, and their evaluation in shellfish tissue as standard techniques for the detection of domoic acid, as a model toxin compound. The key advantages in the use of these biosensor techniques for screening hybridomas in monoclonal antibody production were the real time observation of molecular interaction and rapid turnaround time in analysis compared to enzyme immunoassays. The multichannel prototype instrument was superior with 96 analyses completed in 2h compared to 12h for the single channel and over 24h for the ELISA immunoassay. Antibodies of high sensitivity, IC50's ranging from 4.8 to 6.9ng/mL for monoclonal and 2.3-6.0ng/mL for polyclonal, for the detection of domoic acid in a 1min analysis time were selected. Although there is a progression for biosensor technology towards low cost, multiplexed portable diagnostics for the food industry, there remains a place for laboratory-based SPR instrumentation for antibody development for food diagnostics as shown herein.


Assuntos
Análise de Alimentos/métodos , Ácido Caínico/análogos & derivados , Toxinas Marinhas/análise , Frutos do Mar/análise , Ressonância de Plasmônio de Superfície/métodos , Animais , Anticorpos Imobilizados/química , Anticorpos Monoclonais/química , Desenho de Equipamento , Análise de Alimentos/instrumentação , Hibridomas , Imunoensaio/instrumentação , Imunoensaio/métodos , Ácido Caínico/análise , Limite de Detecção , Camundongos Endogâmicos BALB C , Coelhos , Ressonância de Plasmônio de Superfície/instrumentação
2.
Artigo em Inglês | MEDLINE | ID: mdl-22035229

RESUMO

Carbon black (C.I. 77266) is an insoluble pigment produced by the partial combustion of hydrocarbons. The pigment is known by several synonyms, including vegetable carbon, lamp black and carbon ash, that correspond to the raw materials and methods used for its production. Vegetable carbon (E153) is permitted for use in colouring food in the European Union. The US Food and Drug Administration (USFDA) has not approved the use of any type of carbon black for colouring food, although the agency batch certifies the pigment as D&C Black No. 2 for use in colouring certain cosmetics. Since carbon black (as vegetable carbon) may be present in food products offered for import into the United States, the USFDA's district laboratories need a qualitative analytical method for determining its presence. We have developed an extraction method for this purpose. A sample is broken down and dissolved with nitric acid. The resulting solution is filtered and treated with hydrochloric acid to dissolve any black iron oxide also present as a colour additive. A black residue remaining on the filter paper indicates the presence of carbon black in the food. We confirmed the presence of carbon black in residues from several standards and food products using Raman spectroscopy. The limit of detection for this method is 0.0001%.


Assuntos
Análise de Alimentos , Fuligem/química , União Europeia , Padrões de Referência , Análise Espectral Raman , Estados Unidos , United States Food and Drug Administration
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