Slide 1: How are the in vitro GARD assays used in real life?
How are the in vitro GARDTM assays used in real life? Cases and applications for testing of skin and respiratory sensitizers Andy Forreryd, PhD Copyright Protected and all rights reserved. This is the property of SenzaGen and must not be reproduced in part or full without SenzaGen’s written permissi
Slide 2: About SenzaGen
About SenzaGen GARDTM • Founded in 2010 • Business model: global Our lead product, GARDTM stands • Spin-out from Lund University industrial and CRO partnerships for Genomic Allergen Rapid after over 10 years research • Own laboratory, continuous Detection and is a state-of-the art • Highly multidisc
Slide 3: The GARD platform – Currently available assays
The GARD platform – Currently available assays GARD™skin (200 genes) To identify the skin sensitization hazard of chemicals GARD™skin Medical Device (200 genes) To identify the skin sensitization hazard of medical devices GARD™potency (51 genes) Skin sensitization potency classification according to
Slide 4: The GARD platform – One testing platform for many answers
The GARD platform – One testing platform for many answers GARDskin Medical Device Copyright Protected and all rights reserved. This is the property of SenzaGen and must not be reproduced in part or full without SenzaGen’s written permission.
Slide 5: The GARD technology platform
The GARD technology platform Human relevant cells in combination with Genomics and machine learning Copyright Protected and all rights reserved. This is the property of SenzaGen and must not be reproduced in part or full without SenzaGen’s written permission.
Slide 6: The GARD platform – how it works
The GARD platform – how it works SenzaCells: a human dendritic-like cell-line Skin sensitizer Cellular responses Respiratory Sensitizer Non-sensitizer Copyright Protected and all rights reserved. This is the property of SenzaGen and must not be reproduced in part or full without SenzaGen’s written p
Slide 7: The GARD platform – how it works
The GARD platform – how it works Cellular response is monitored using biomarker signatures – Not only a single biomarker GARDskin prediction signature 200 genes Recognition of foreign substances e.g. TLRs, RXR, AHR Immunological self-defence mechanisms e.g. CD80, CD86 Cellular stress responses e.g.
Slide 8: The GARD platform – how it works
The GARD platform – how it works Keratinocytes Dendritic cells T-cells Key Event 1 Key Event 2 Key Event 3 Key Event 4 >Pro-inflammatory >Antigen recognition Covalent binding to signalling >DC activation Activation/ proteins >Cytoprotective >DC maturation proliferation of T-cell signalling >DC migra
Slide 9: How to GARD your product in steps
How to GARD your product - in 6 Steps Copyright Protected and all rights reserved. This is the property of SenzaGen and must not be reproduced in part or full without SenzaGen’s written permission.
Slide 10: GARDskin – Performance data
GARDskin – Performance data The standard assay protocol is applicable Data set for small molecules Sensitivity Specificity Accuracy Reference in general: from cosmetics ingredientsoftothe Validation various GARD industrial assay 93% (51/55) 96% (24/25) 94% (75/80) Johansson, 2019 chemicals, includin
Slide 11: GARDskin & GARDpotency – REACH registration
GARDskin & GARDpotency – REACH registration “The REACH Regulation allows the use of non-adopted in vitro methods in case they meet the EURL ECVAM criteria for entering pre-validation. For the GARD assay this criteria is met, as it is currently being validated.” “The current REACH information require
Slide 12: GARDskin – Broad applicability domain
GARDskin – Broad applicability domain 100 The standard assay protocol is Mean accuracy (89%) applicable for small molecules 80 in general: from cosmetics Active Pharmaceutical Ingredients 37 ingredients to various industrial Disinfectants chemicals, including pre- and pro- Dyes haptens Flavoring age
Slide 13: More info at: www.senzagen.com
More info at: www.senzagen.com Copyright Protected and all rights reserved. This is the property of SenzaGen and must not be reproduced in part or full without SenzaGen’s written permission.
Slide 14: Case study I – GARDskin for difficult to test substances
Case study I – GARDskin for ”difficult to test” substances Why substances can be difficult to test: Pre/Pro haptens: Activation may be needed to create the allergen. SenzaCells: Aldehyde dehydrogenases (ALDH) Cytochrome p-450 (CYP) N-acetyltransferase 1 (NAT-1) Solubility: High log Po/w value or oth
Slide 15: Case study I – GARDskin for difficult to test substances
Case study I – GARDskin for ”difficult to test” substances Compound Pre/pro High DPRA ARE-NRF2 h-CLAT GARD hapten logP (TG442C) (TG 442D) (TG442E) (TGP 4.106) 2-Aminophenol YES - S S S S1 2-nitro-1,4-Phenylendiamine YES - S S S S1 Aniline YES - NS NS S S1 Cinnamic alcohol YES - S S S S1 Ethylene dia
Slide 16: Case study II – Testing of UVCBs
Case study II – Testing of UVCBs UVCBs: Unknown or Variable The challenges: Complexity and uncertainty of composition, Complex reaction compositions; Hydrophobic, often has solubility issues for products and Biological materials water-based test systems Vehicle: DMSO Vehicle: H2O Vehicle: DMF/Glycer
Slide 17: Case study II – Testing of UVCBs
Case study II – Testing of UVCBs Materials & Methods Test item GARDskin Additional solubility 7 UVCBs with known tests performed to average MW select suitable solvents Conclusion Results in comparison with existing in vivo data GARDskin: good applicability potential Test Existing data Existing GARDs
Slide 18: Case study III – Testing of natural mixtures
Case study III– Testing of natural mixtures Henna: natural dye from Lawsonia inermis. Henna-based hair colouring products: often considered safer alternatives to synthetic hair dyes containing known skin sensitizers such as p-phenylenediamine (PPD). However, skin allergic reactions to henna products
Slide 19: Case study III – Testing of natural mixtures
Case study III– Testing of natural mixtures Table 1.GARDskin prediction results for commonly used hair dye ingredients Human Animal Test materials mDPRA IL-18 USENSTM GARDTM skin classification classification Reference controls Dimethyl sulfoxide NS NS NS NS NS NS Glycerol NS NS NS NS NS NS Sodium d
Slide 20: Case study III – Testing of natural mixtures
Case study III– Testing of natural mixtures Table 2. GARDskin prediction results for 10 Conclusion commercial henna-based products Henna PPD PPD mDPRA IL-18 USENS™ GARDskin™ GARDskin show high Product (label) concordance to human data 1 1.091 ± S S S S for testing of hair dye 0.028 2.970 ± ingredien
Slide 21: Case study IV – Testing of oil-based mixtures
Case study IV– Testing of oil-based mixtures The Customer: a leading supplier of natural based functional ingredients/ raw materials for cosmetic and personal care applications The problem: An oil-based mixture with unpleasant and characteristic smell • Used as raw materials sold to cosmetic and per
Slide 22: Case study IV – Testing of oil-based mixtures
Case study IV– Testing of oil-based mixtures Methods & Results (GARDskin) Test item 1 Test item 2 Test item 3 Oil mixture containing Distilled oil mixture containing Pure chemical x 1200 ppm chemical x 10 ppm chemical x Non sensitizer Non sensitizer Sensitizer GARDskin Decision Values (DVs) 1 Conclu
Slide 23: Case study V – Testing Medical Devices for skin sensitization hazard
Case study V– Testing Medical Devices for skin sensitization hazard In vitro skin sensitization testing in the Biological Evaluation of Medical Devices according to ISO 10993-12:2012 • Use polar and non-polar extraction vehicles according to ISO 10993-12:2012 (saline, sesame oil, super refined olive
Slide 24: Case study V – Testing Medical Devices for skin sensitization hazard
Case study V– Testing Medical Devices for skin sensitization hazard Extraction steps of solid material follow the international standards for safety assessment of Medical Device (ISO 10993-12). Saline titration Oil titration 1 1 sensitizer sensitizer 0 Decision Values (DVs) 0 Decision Values (DVs) n
Slide 25: Case study V – Testing Medical Devices for skin sensitization hazard
Case study V– Testing Medical Devices for skin sensitization hazard Copyright Protected and all rights reserved. This is the property of SenzaGen and must not be reproduced in part or full without SenzaGen’s written permission.
Slide 26: Case study VI – Safety assessment of e-liquids
Case study VI – Safety assessment of e-liquids Materials & Methods In this customer study, GARDskin, GARDpotency and GARDair were used to predict and compare the skin and respiratory sensitisation potential of three experimental and two commercial e-liquids. To our knowledge this is the first publis
Slide 27: Case study VI – Safety assessment of e-liquids
Case study VI – Safety assessment of e-liquids The typical composition of e-liquids Base liquid Propylene Glycol (PG) and/or (aerosol former) Vegetable Glycerol (VG) E-liquids Flavorings Natural and/or artificial Nicotine (with or without) E-liquids are usually complex mixtures, especially when they
Slide 28: Case study VI – Safety assessment of e-liquids
Case study VI – Safety assessment of e-liquids GARDair prediction A controls: 5 out of 7 accurately classified as respiratory sensitizers, no false positives. Sensitivity 71%, Specificity 100%, Overall accuracy 89% B test substances: None of the base liquids nor commercial e-liquids were classified
Slide 29: Case study VI – Safety assessment of e-liquids
Case study VI – Safety assessment of e-liquids GARDskin prediction : GARDpotency prediction : 2 commercially available flavoured Test substances: 2 commercially available flavoured e-liquids e-liquids were predicted as skin were further classified as weak skin sensitizers (1B) sensitizers Copyright
Slide 30: Case study VI – Safety assessment of e-liquids
Case study VI – Safety assessment of e-liquids Conclusion The GARD platform offers an integrated test strategy for assessment of skin and respiratory sensitization potential of complex mixtures such as e-liquids. The assays are potentially useful for product development and hazard identification of
Slide 31: The GARD platform – How can I get my substances tested?
The GARD platform – How can I get my substances tested? Contact us: We help to design the testing strategy for your specific substances. Select assay(s): GARDskin, GARDpotency, GARDair, GARDskin Medical Device Test Substance Questionnaire: Do you have information on vehicles? If not, we evaluate it
Slide 32: The GARD platform – Where can I get my testing done?
The GARD platform – Where can I get my testing done? Licence Labs*: Burleson Research Technologies Eurofins BPT MB Research Laboratories Distributors*: Charles River Laboratories Eurosafe Guangzhou CHN-ALT Biotech Co., Ltd PKDerm Woo Jung BSC XCellR8 * In alphabetic order Copyright Protected and all
Slide 33: Information and contacts
Information and contacts www.senzagen.com Andy Forreryd, PhD Joshua Schmidt, PhD Key Account Manager & Scientific Liaison Business Development Director Americas andy.forreryd@senzagen.com Joshua.schmidt@senzagen.com +46 734331777 +1 651.440.5691 Copyright Protected and all rights reserved. This is t