Lorem Ipsum Lorem. South African Society of Dairy Technology Symposium 24-26/04

Transcription

1 Lorem Ipsum Lorem South African Society of Dairy Technology Symposium 24-26/04

2 UNISENSOR Unisensor is a Belgian company researching, developing and manufacturing diagnostic tools for the detection of contaminants present in food products.

3 Company Timeline milestones Twinsensor PATENTED R&D EFFORT EU RESEARCH PROGRAMS GOOD FOOD BEADYPLEX EXTENSO CONCEPT CONFFIDENCE REVOLUTIONARY MILK ANALYSIS PLATFORM 014

4 X 17 MULTIPLE: molecules in a single test Detection level at EU MRL

5 Our Product Development Strategy In two words 1 x 17 x MULTIPLEXING + CONTAMINANTS Multiple tests in one (antibiotics) Reliable & Usable everywhere Rapid & Cost-effective Validated by external parties 05

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7 Scheme of analytical methods for antibiotic residues analysis Complexity Price TESTING PRINCIPLES Confirmation METHODS AVAILABLE Composition PHYSICO CHEMICAL LC-MS Structure IMMUNOAssays ELISA LFIA Activity MICROBIOLOGICAL Inhibition Screening 07

8 Unmet needs for a screening test 1. Multiple families ID in a single test (vs ELISA) 2. Sensitive for all/most residues (vs microbiological methods) 3. High Throughput (+100 samples/day) 4. Cost-effective and time-saving Adding Multi-Matrix dimension 08

9 - All-In-One screening Flow Cytometric Immunoassays Assay principle Immunoanalysis: detection of interaction events (receptor & antibodies). Flow Cytometry: multi-parametric characterization of microparticles (beads) by lazer. Multiplexing via microparticles characterized by: 1. Different sizes 2. Different Internal fluorescence intensities 09

10 - All-In-One screening Flow Cytometric Immunoassays Assay principle FCIA: simultaneous detection of residues of antibiotics from 10 families in one sample in one analysis 10 different ELISA kits combined in one test 010

11 - All-In-One screening Flow Cytometric Immunoassays Assay principle FLOW CYTOMETRIC COMPETITIVE IMMUNOASSAY 011

12 - All-In-One screening Flow Cytometric Immunoassays Assay principle Determination of assay signal intensity BL RL Classification of particles Size Internal fluorescence 012

13 Workflow for Antibiotic residues 1 2 Sample preparation Immunoassay Automatic Reading 3 4 Automatic Reporting Flow cytometer (NovoCyte 2000, ACEA Biosciences)

14 Analytical performance Validation Following CRLs Guidelines for the validation of Screening methods for residues of veterinary medicines (20/01/2010) based on Commission Decision 2002/657/EC Specificity/selectivity Cut-Off values Detection capabilities False positive rates Robustness Applicability 014

15 Analytical performance Detection Limits (CCβ) CCβ (β error 5%) concentration for each antibiotic recognized by the FCIA for which at least 95% of the samples fortified at this level are identified as screened positive according to the established assay cut-off values CCβ determined for all detected molecules Most of CCβ MRL 038

16 Analytical performance Detection Limits (CCβ) Profile of antibiotic detection (non exhaustive list) 039

17 Analytical performance False positive rate Bead n Blank samples n Screen positive False positive rate (%) AMINOGLYCOSIDES_A AMINOGLYCOSIDES_B BETA-LACTAMS LINCOSAMIDES COLISTIN MACROLIDES_A MACROLIDES_B False positive rate < to those usually atributed to other screening methods for antibiotics SULFONAMIDES PHENICOLS TETRACYCLINES (F)QUINOLONES PLEUROMUTILINES ,6 040

18 Analytical performance Robustness Parameter Condition 1 (control) Condition 2 Condition 3 SAMPLE EXTRACTION ASSAY PROCEDURE EXTRACTION VOLUME 1 ML 0,8 ML 1,2 ML AGITATION EXTRACTION TIME 10 MINUTES 5 MINUTES 30 MINUTES AGITATION SPEED 6,5 AGITELEC 4,5 AGITELEC 2,5 AGITELEC SAMPLE EXTRACTS STORAGE EXTRACTION + ANALYSIS 4 C ON -20 C ON EXTRACTION METHOD AGITATION + CENTRIFUGATION STOMACHER - NUMBER OF PLATE WASHING STEPS TEST INCUBATION TEMPERATURE RT 4 C 37 C INCUBATION TIME COMPETITIVE STEP 30 MINUTES 15 MINUTES 60 MINUTES INCUBATION TIME REPORTERS STEP 15 MINUTES 5 MINUTES 30 MINUTES No impact of any of the evaluated parameters on assay performance Extraction and analysis need to be performed the same day Very robust method PLATE READING DELAY TEST + READING 1H DELAY ON DELAY REAGENTS (BEADS AND PRIMARY BINDERS) BATCH 1 BATCH 2 BATCH 3 OTHER OPERATOR A B C CYTOMETERS NOVOCYTE ACEA ACCURI C6 BD - REAGENTS STABILITY SAME DAY 3 MONTHS 4 C 6 MONTHS 4 C 041

19 Analytical performance Applicability Porcine muscle Bovine muscle Poultry muscle Fish (salmon and colin) Milk Applicable to Honey Eggs FLEXIBILITY Extraction procedure adaptation for different matrices Cut-off modification for modulation of applicability or sensitivity 019

20 Analytical performance - Conclusions 020

21 Analytical performance - Conclusions 021

22 Developments Mycotoxines Foodborne pathogenic bacteria Antibiotics residues Hormones Sedatives Antiinflammatory drugs Food allergens 022

23 Allergens

24 Flow Cytometric Immunoassays Assay principle FLOW CYTOMETRIC SANDWICH IMMUNOASSAY 024

25 09 BEADYPLEX Flow Cytometric Immunoassays Assay principle Determination of assay signal intensity BL RL B Classification of particles 025

26 LOD/LOQ Range BLG 0, ppm Casein 0, ppm Egg 1,25 80 ppm Peanut 1,25 80 ppm Soy 1,25 80 ppm 026

27 Hormones Sedatives Anti-inflammatory drugs

28 Hormones Sedatives Anti-inflammatory drugs Screening multi-class and multi-residue of veterinary drugs in meat/milk or urine by FCIA Growth promoters Post-mortem test Ante-mortem test Corticosteroids 028

29 Thank you for your attention FCIA team Alessandro Gallo Alexandre Monteforte Cédric Pety de Thozée Celia Suárez Pantaleón