Smart micro-sensing: antibodies and aptamer-based micro-elisa as performing off line/on line tool for allergens and mycotoxins detection in foods

Transcription

1 Smart micro-sensing: antibodies and aptamer-based micro-elisa as performing off line/on line tool for allergens and mycotoxins detection in foods L. Boschis, N. Ciprianetti, Elena Gulì, D. Spadaro, M.L. Gullino, R. Rinaldi, C. Garino, M. Arlorio 1 1

2 FDM FOOD DIGITAL MONITORING* a new control model for food production and processing based on the Process Analytical Technology, Internet of Things, open data and 'big data'. RESEARCH CENTERS TECH COMPANIES END USER COMPANIES *funded with the contribution of the Piedmont Region with European Funds for Regional Development 2

3 FDM FOOD DIGITAL MONITORING* The food smart factory *funded with the contribution of the Piedmont Region with European Funds for Regional Development 3

4 FDM FOOD DIGITAL MONITORING* Transformation 1 Transformation 2 Transformation n Raw material analysis 1 analysis 2 analysis 3 analysis n Final product micro-elisa Data storage Data analysis and visualization *funded with the contribution of the Piedmont Region with European Funds for Regional Development 4

5 An off line/on line tool for food quality control easy as a lateral flow precise as laboratory ELISA 5

6 It is composed by Disposable and ready-to-use Lab on Chip (LOC) cards which are the sensitive element of the device A portable, easy to use and automated reader able to display results and connect to smartphone, tablet or computers. 6

7 THE LAB-ON-CHIP CARD Microfluidics enable miniaturization and automation The Lab-on-Chip card has 4 reaction lines and all the reagents are pre-filled as ready to use 7

8 THE MICROFLUIDIC CIRCUIT IS DESIGNED TO EXECUTE ELISA OR ELASA PROTOCOLS ELISA ELASA Non Competitive ELISA Competitive ELISA Indirect ELISA 8

9 THE READER a complex set of valves and pumps enable the movement of reagents inside the card 9

10 DETECTION METHOD 450 nm or 630 nm Detection chamber light source photodiode 10 9

11 Tensione fotodiodo (mv) Concentrazioni standard (ppt) 11

12 CARCIROGENIC FOOD CONTAMINANT AND FOOD ALLERGEN ANALYSIS HAVE BEEN TESTED Aflatoxin B1 carcirogenic mycotoxin Lysozyme and Gliadin allergens 12 10

13 Absorbance (OD) LYSOZYME (example of non competitive assay) standard curve Lysozime 2,5 2 1,5 1 0,5 MicroElisa 20 µl Commercial Kit Session CV% ,5 6,25 12,5 ng/ml A = -log (I/I₀) 13

14 B/B0 AFLATOXIN B1 (example of competitive assay) standard curve MicroELISA 20 μl Commercial Kit Aflatoxin B1 Session CV% ,01 0, ng/ml A = -log (I/I₀) 14

15 AFLATOXIN B1 (example of competitive assay) DIFFERENT REAGENT CONCENTRATIONS MODULATE THE ANALYTICAL CURVE 15

16 All immunoassays can be performed by : In Biological fluids and Food (Rough or processed samples, according to protocols): Hormones Contaminants Toxins Allergens Pathogens Water Milk Food 16

17 APTAMERS micro-sensing Easier and more economical to produce Aptamers are capable of grater specificity and affinity than antibodies Aptamers can easily be modified chemically to tailored properties Aptamer are much more stable at ambient temperature than antibodies 17

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