Allergen testing at Eurofins

A Short Introduction

Dr Koh Yew Ming, FMIC, MJMM

BD General Manager, Malaysia & Indonesia
M: +6012 482 6123
yewmingkoh@eurofins.com

06.02.2021

xxxxxx www.eurofins.de
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About Eurofins Scientific

As a world leader and innovator in analytical testing, Eurofins is the quiet hero that stands between you and the hazards of an ever-
changing, highly complex world. Our continuous innovation and ceaseless vigilance mean you can trust the products you consume, the
medicines you depend on and the environment you live in. We walk beside you every day, keeping your family safe, making our world
better.

Day in, day out, we are testing for life.

2016
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Environment BioPharmaceuticals
N°1 worldwide Top-10 worldwide

“Recent trends in global food production, processing, distribution

and preparation are creating an increasing demand for food safety
research in order to ensure a safer global food supply.”
World Health Organisation
2016
Food & Feed Testing is one of Eurofins’ 4 large business
lines. Eurofins is #1 worldwide, as well as in Europe,
USA, and many major economies

Testing for Food & Feed Environment Clinical

Pharma/Biotech/Agrosciences Testing Testing Diagnostics

N°1 to N°3* N°1* N°1* Start-Up

Eurofins position worldwide worldwide worldwide Start 2014
Start 2000-2005 Start 1987 Start 2000

N° 1 Worldwide in Pharma N° 1* worldwide N° 1* Worldwide Establishing leadership in

Products Testing N° 1 in the USA N° 1 in the USA targeted higher-growth
Eurofins ranking

N° 1 Worldwide in Discovery N° 1 in Europe N° 1 in Europe innovative niche areas

Pharmacology Services N° 1 in Germany N° 1 in Germany (e.g. genomics, infectious
N° 1 Worldwide in Agroscience N° 1 in France diseases, etc.) of the
N° 1 in France
CRO services clinical testing market,
N° 1 in Nordics/Scandinavia N° 1 in Nordics/Scandinavia
Among top 5 global providers of mainly in the U.S. and
N° 1 in Benelux N° 1 in Ireland
central laboratory and genomic Europe, as well as market
N° 1 in the UK & Ireland N° 1 in Benelux access through local
services N° 1 in Spain N° 2 in Spain laboratories in many large
N° 1 or 2 in most segments/
N° 1 in Brazil N° 2 in Japan markets worldwide to
countries in Europe and the USA
N° 1 in Agro Testing EU distribute advanced tests.4

Title: The Potential of Testing to Protect the Environment and the Health of All Document Name: AOAC_Key note speech_202008_v4_final.pptx Document owner: Sabine Nitsch Last modified on: 27/08/2020 4
 Eurofins Laboratories Worldwide

Employees Laboratories Countries Eurofins

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 Eurofins presence in Asia (excl. IT)
India: Food Japan:
 Food (x2) Environment  Environment (x4)
Pharma
 Environment Consumer Products  Asbestos
 Agro sciences Genomics  Food
 Pharma Clinical Diagnostics  Pharma
Agro science
 Genomics Materials  Consumer Products
 Clinical Genetics  Genomics
 Consumer Products (x2)  Clinical Genetics
 Agro sciences
Middle East  Material Sciences
 Clinical Diagnostics
South Korea:
Malaysia:  Environment
 Environment  Cosmetics
 Consumer Products  Electrical & Electronic
 Rubber  Food
 Food
China:
Singapore:  Food (x5)
 Environment
Taiwan:
 Consumer Products
 Food
 Food  Electrical & Electronic
Vietnam:
 Bunker Oil
 Environment  Material Sciences
 Food (x3)
 Clinical trials
 Drug Discovery  Clinical trials
 Consumer Products
 Technologies
 Electrical &
 Clinical Diagnostics
Electronic
Hong Kong:
Thailand:
 Material Sciences  Food
 Electrical & Electronic
 Digital
 Agro sciences
 Consumer Products
 Food Philippines:
 Food
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9 Food testing: Eurofins serves the complete
“Food Chain”

Eurofins Malaysia 32
233
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u Eurofins’ Food & Feed testing offering is the most
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r comprehensive in the market
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s Agricultural
M production, product Production Retail & Distribution
a development
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s Sensorial Dioxins Veterinary drug Vitamins Quality Control
Microbiology
i residues

Authenticity Pesticides Organic residues POPs Labelling GMO

Allergens Heavy metals Irradiation Nutritional Purity

Mycotoxins

Testing offer available either locally or through Eurofins Competence Centres

Food Allergy and Celiac Disease

FOOD ALLERGY CELIAC DISEASE

Food allergy is an abnormal Celiac disease is an autoimmune

response to a foodstuff, triggered by disease that occurs where the
the body's immune system. ingestion of gluten leads to
damage in the small intestine.

It is estimated that celiac disease affect 1% people worldwide;

and approximately 2-5% of adults and 4-10% of children are affected by food
allergies.

The global food allergen testing market is driven by the increasing number
of food allergies reported to medical establishments

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The Importance of Allergen Testing

Currently, the only treatment for

food allergies or celiac disease
is lifelong adherence to a strict Identification
allergen-free diet. of food
ingredients
Food allergens may be present
as an ingredient but also Allergen-
inadvertent via cross
contamination when sharing management
food processing machines.
Potential
cross-
This implies a traceability of the contamination
food production process, from
the raw material to the final
product.

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Allergens management

An effective allergen management system covers all aspects of the food product
supply chain from sourcing raw materials through manufacturing and packaging,
to the finished product.

• Identifying and controlling allergens

• Cleaning the facility, equipment, and tools to prevent allergen cross contact
Monitoring by surface (swab) or rinse water testing

Raw product Production Retail

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 Our service

• Analysis of allergens

Cereals containing
Crustaceans Peanuts Eggs Molluscs
gluten

Soybeans Sulfite (limit 10 ppm) Sesame Celery Mustard

Fish Milk incl. lactose Lupine „Treenuts“ Coconut

• Wide range of methods for allergen detection

• Assistance in the selection of appropriate methods
• Consultancy and assistance in risk assessment

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 Competence Center: Food Allergens

• Many years of experience in the field of allergen analysis

• Fast service & high capacities
• Using the latest scientific methods and method development
• Close cooperation with certification bodies, methods suppliers and
international partners

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Allergen Testing - Requirements

 Highly specific
Differentiation between closely related species, like celery and parsley

 Low detection limits

sensible people react at very low doses (low ppm range)

 Suitable for diverse matrices and

processing stages

 Reasonable testing
of high sample numbers

 Quantification

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Detection methods

ELISA

Relevance in routine testing

 Immunological method
(specific antibody recognises protein)

PCR
 Molecular biological method
(detection of specific DNA)

LC-MS/MS
 Mass spectrometry
(direct detection of peptides)

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ELISA (Enzyme-linked Immunosorbent Assay)

Principle Sandwich ELISA

 1st antibody bound at surface

 Addition of sample extract (with allergen)

1st antibody binds allergen 2nd antibody

 2nd antibody binds allergen,

formation of „Sandwich“
2nd antibody connected with enzyme

 Addition of substrate 1st antibody

 Enzyme at 2nd antibody converts substrate

into a colour
– Quantification due to standards with
known amount of allergen

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Advantages ELISA

 Detection of protein (allergen = protein)

 fast

 quantitative

 high sensitvity

(LoD of 0.2 mg protein)

 „ready-to-use“ kits for almost all allergens

 Automatisation possible

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 Standardization with Eurofins ELISA Kits

SENSISpec Allergen ELISA

• Comprehensive list of Allergen ELISA Kits covering all relevant food allergens
(including a Swabbing Kit)

SENSISpec Allergen Detection ELISA Kits

Allergen ELISA Kits provided by

Eurofins Technologies
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 Allergens SENSISpec ELISA

ELISA kits ELISA kits

SENSISpec INgezim Gluten R5 SENSISpec ELISA Macadamia nut
SENSISpec INgezim Gluten R5 (Hydrolyzed samples) SENSISpec ELISA Milk
SENSISpec ELISA Almond SENSISpec ELISA Molluscs (Tropomyosin)
SENSISpec ELISA Beta-Lactoglobulin SENSISpec ELISA Mustard
SENSISpec ELISA Casein SENSISpec ELISA Ovalbumin
SENSISpec ELISA Cashew SENSISpec ELISA Brazil Nut
SENSISpec ELISA Coconut SENSISpec ELISA Peanut
SENSISpec ELISA Crustaceans (Tropomyosin) SENSISpec ELISA Pecan Nut
SENSISpec ELISA Egg White SENSISpec ELISA Pistachio
SENSISpec ELISA Fish (Parvalbumin) SENSISpec ELISA Sesame
SENSISpec ELISA Hazelnut SENSISpec ELISA Soy
SENSISpec ELISA Lupin SENSISpec ELISA Total Soy Protein
SENSISpec ELISA Lysozyme SENSISpec ELISA Walnut

Company Presentation Eurofins Food Germany

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PCR (Polymerase Chain Reaction)

 DNA Extraction
whole DNA of the sample

 Amplification of a specific DNA region

e.g. part of a specific peanut gene

 Detection of the amplified DNA

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Advantages PCR

 fast

 sensitive [1-50 pg DNA / 1-40 mg allergenic food]

 Database with a vast amount of DNA information

+ many PCR kits available

 high sample numbers

 Quantification + multiplexing possible

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 (Liquid Chromatography
LC-MS/MS
Tandem Mass spectrometry)

Principle:

- Protein extraction + digestion

- Chromatographic separation (LC)
- MS/MS ionisation + detection

MS/MS

enzymatic LC
digestion fragmentation
seperation
PROT
Peptides precursor fragment
Ions (fixed) Ions (fixed)

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Advantages LC-MS/MS

Multi-Screening
 Several allergens at the same time

Highly specific
 8 data points per peptide
usually 4 peptides per peptide

Highly reliable
 If one peptid is instable or not detectable, the other 2 or 3 are still detectable

Well suitable for processed matrices

 LC-MS/MS detection bases on primary structure of the protein/peptide
therefore better suitable for processed products

Sensitive
 Comparable LoDs

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SENSIStrips Allergen LFD – First wave!

4 kits available from December 2020

Peanut Almond Crustacean Mustard

No. of Sensitivity Sensitivity Sensitivity

Article no. LFD kits
Tests Food Matrix Swabbing Rinse water

HU0030128 SENSIStrip Peanut 20 2.5 ppm 0.07 µg/cm2 0.17 mg/L

HU0030110 SENSIStrip Almond 20 2.5 ppm 0.07 µg/cm2 0.17 mg/L

SENSIStrip Crustaceans
HU0030115 20 25 ppb 2 ng/cm2 1.7 µg/L
(Tropomyosin)

HU0030125 SENSIStrip Mustard 20 2.5 ppm 0.07 µg/cm2 0.17 mg/L

Shelf life: 12 months (stored between 2ºC and 8ºC)

SENSIStrip LFD Product Line

• Kits provided with all the necessary material

to perform 20 individual analysis per kit.
• Easy to use lateral flow format, giving a
rapid and accurate “on site” test
• High sensitivity and specificity, thanks to
the combination of biotinylated antibodies and
gold nanoparticles conjugated to streptavidin
• Possibility of semi-quantitative results
• Low pricing
• Commercial and Technical support
• Orders: technologies.hu@eurofins.com
 Thank you for your attention

We are looking forward to pursue and

expand our cooperation with you!

Contact Us !
MY01-BMU.NDSC@eurofins.com

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