Lorem Ipsum Lorem South African Society of Dairy Technology - PowerPoint PPT Presentation

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

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

Company Timeline milestones 2005 2015 2017 R&D EFFORT Twinsensor EU RESEARCH PROGRAMS EXTENSO BEADYPLEX “ GOOD FOOD ” PATENTED “ CONFFIDENCE ” CONCEPT REVOLUTIONARY MILK ANALYSIS PLATFORM page 014

X 17 MULTIPLE: + 100 molecules in a single test Detection level at EU MRL

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

BEADYPLEX Scheme of analytical methods for antibiotic residues analysis Complexity Confirmation T ESTING M ETHODS P RINCIPLES AVAILABLE Price LC-MS Composition PHYSICO CHEMICAL ELISA Structure IMMUNOAssays LFIA MICROBIOLOGICAL Inhibition Activity Screening page 07

BEADYPLEX 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 page 08

BEADYPLEX - “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 page 09

BEADYPLEX - “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 page 010

BEADYPLEX - “All -In- One” screening Flow Cytometric Immunoassays – Assay principle FLOW CYTOMETRIC COMPETITIVE IMMUNOASSAY page 011

BEADYPLEX - “All -In- One” screening Flow Cytometric Immunoassays – Assay principle Determination of assay signal intensity BL RL Classification of particles Size Internal page fluorescence 012

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

BEADYPLEX 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 page 014

BEADYPLEX Analytical performance Detection Limits (CC β ) CC β ( 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 ✓ CC β determined for all detected molecules ✓ Most of CC β ≤ MRL page 038

BEADYPLEX Analytical performance Detection Limits (CC β ) Profile of antibiotic detection (non exhaustive list) page 039

BEADYPLEX Analytical performance False positive rate n Screen False positive Bead n Blank samples positive rate (%) A MINOGLYCOSIDES _A 117 0 0 A MINOGLYCOSIDES _B 117 0 0 B ETA - LACTAMS 107 0 0 L INCOSAMIDES 123 0 0 False positive rate < to those C OLISTIN 122 0 0 usually atributed to other M ACROLIDES _A 113 0 0 screening methods for antibiotics M ACROLIDES _B 113 0 0 S ULFONAMIDES 107 0 0 P HENICOLS 118 0 0 T ETRACYCLINES 117 0 0 (F)Q UINOLONES 110 0 0 P LEUROMUTILINES 122 2 1,6 page 040

BEADYPLEX Analytical performance Robustness Parameter Condition 1 (control) Condition 2 Condition 3 E XTRACTION VOLUME 1 M L 0,8 M L 1,2 M L ✓ No impact of any of the evaluated parameters on A GITATION EXTRACTION TIME 10 MINUTES 5 MINUTES 30 MINUTES assay performance S AMPLE A GITATION SPEED 6,5 A GITELEC 4,5 A GITELEC 2,5 A GITELEC EXTRACTION ✓ Extraction and analysis S AMPLE EXTRACTS STORAGE E XTRACTION + ANALYSIS 4°C ON -20°C ON need to be performed E XTRACTION METHOD A GITATION + CENTRIFUGATION S TOMACHER - the same day N UMBER OF PLATE WASHING STEPS 3 1 2 ✓ Very robust method T EST INCUBATION TEMPERATURE RT 4°C 37°C A SSAY I NCUBATION TIME COMPETITIVE STEP 30 MINUTES 15 MINUTES 60 MINUTES PROCEDURE I NCUBATION TIME REPORTERS STEP 15 MINUTES 5 MINUTES 30 MINUTES P LATE READING DELAY T EST + READING 1 H DELAY ON DELAY R EAGENTS ( BEADS AND PRIMARY B ATCH 1 B ATCH 2 B ATCH 3 BINDERS ) O PERATOR A B C O THER page C YTOMETERS NOVOCYTE ACEA A CCURI C6 BD - 041 R EAGENTS STABILITY SAME DAY 3 MONTHS 4°C 6 MONTHS 4°C

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

BEADYPLEX Analytical performance - Conclusions page 020

BEADYPLEX Analytical performance - Conclusions page 021

BEADYPLEX Developments Mycotoxines Foodborne pathogenic bacteria Antibiotics residues Food Hormones allergens Sedatives Anti- inflammatory drugs page 022

Allergens

BEADYPLEX Flow Cytometric Immunoassays – Assay principle FLOW CYTOMETRIC SANDWICH IMMUNOASSAY page 024

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

BEADYPLEX LOD/LOQ Range 0,625 – 40 ppm BLG 0,625 – 40 ppm Casein 1,25 – 80 ppm Egg 1,25 – 80 ppm Peanut 1,25 – 80 ppm Soy page 026

Hormones Sedatives Anti-inflammatory drugs

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

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