PRE Technologieforum 003 070118 JBE2007/Vortraege/Beator.pdf · Therapie und Studienzentrum für...
Transcript of PRE Technologieforum 003 070118 JBE2007/Vortraege/Beator.pdf · Therapie und Studienzentrum für...
1Protagen AG
Protagen AG
Otto-Hahn-Str. 15
44227 Dortmund
+49-231-9742 6300
Successful Strategy for Systematic Identification of Novel Biomarkers
Technologieforum Diagnostik – Frankfurt, 27./28. Nov. 2006
Dr. Jens Beator
Director Protein Biochips
JB/26.11.2006 Slide: 2
Fact Sheet
Corporation on stocks „Aktiengesellschaft“Legal form:
34Employees:
- Bruker Daltonik GmbH, Bremen - Vivascience AG, Hannover- Serva Electrophoresis GmbH, Heidelberg
Industrial partners:
Otto-Hahn-Str. 15
44227 Dortmund
Germany
Location:
Established: 1997
Protagen AG
2Protagen AG
JB/26.11.2006 Slide: 3
Products & Services - Overview
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Business Strategy
Valu
e / R
even
ues
2000 2004 2008
FeeFee--forfor--serviceservice
and Bioand Bio--ITIT
Protein Biochip Protein Biochip
ProductionProduction
Time
Horizon 1
Horizon 2
Horizon 3
Diagnostics Diagnostics
andand
TherapeuticsTherapeutics
2010
3Protagen AG
> 80 autoimmune disease are currently known
„Autoimmune diseases, with the exception of rheumatoid arthritis
and autoimmune thyroiditis, are individually rare, but together they
affect approximately 5 percent of the population in Western
Countries.“
Davidson & Diamond, 2001, N Engl J Med, Vol. 345, No. 5, 340-350
Biomarkers in Autoimmune Diseases
Commonly accepted autoimmune diseases (AID),
in-vitro diagnosis by autoantibodies (profiles) as “biomarkers”:
Type 1 diabetes, autoimmune thyroiditis, systemic lupus erythematodes,
rheumatoid arthritis, mixed connective tissue disease, Sjögren’s syndrome,
pemphigoid diseases, ….
Autoimmune Diseases – Open Questions
Inciting antigens unknown in many AID:
Juv. idiopathic arthritis, inflammatory bowel disease (Morbus Crohn, Colitis
ulcerosa, …), psoriatic arthritis, …
Autoimmune reactions in important other diseases:
Multiple Sclerosis, Alzheimer, breast cancer, prostate cancer, ….
What are the (auto)antigens?
Which autoantibody profiles are suitable biomarkers?
4Protagen AG
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hEx1 Protein Expression Library
• Derived from human fetal braintissue with largest diversity of expressed genes193.500 human protein expression clones with His-tagsbacterial clones grown on PVDF filtersinducible protein expression
• 37.200 protein expressing clonesreact with anti-His antibody
• 16 000 different gene products available for binding assays
2533 different clusters (multiple clones of same gene)
13550 singletons
Büssow et al. 1998,
Lüking, Gutjahr, Cahill, Lehrach, Protagen & MPI f. Molecular Genetics, unpublished
The Unique Resource for Biomarker Discovery
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MACROarrays:
the only product and the largest human protein diversity for
reproducible, unbiased screening
• hEx1 Protein Expression Library (other libraries available)
• Manufactured using robotic systemsFull traceability & identification of clonesReproducible screening product with 16,000 different expression clones
• Low sample consumption (0.5 ml/sample)
• Large amount of antigen available for antibody binding
Unique Strength of the Technology
5Protagen AG
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Patient and control samples
Screening of 37,000 human expression clones
Generation of a putative
disease specific protein subset
Bioinformatical Analysis
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Protein 1 Protein 2 Protein 3 Protein 4 Protein 5 Protein 5 Protein 6 Protein 7 Protein 8 Protein 9
Putative auto-antigens
Facto
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Factor19
Factor20
Factor21
Factor22
Factor23
Factor24
Factor1
Factor2
Factor3
Factor4
Factor5
Factor6
Factor7
Factor8
Factor9
Factor10
Factor11
Bioinformatical analysisIndication-specificmultiplex biochip
Identification of Biomarkers - Strategy
Patient
Sample
Control
Sample
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MACROarray Serum Screening: Alopecia areata
Identification of 23 protein expressing clones,
detected by antibodies in two or more of the five patient pools,
not recognized by serum from 11 controls.
AP
6Protagen AG
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1 21 and 2 show the results of two separate screens of the same
patient, screened separately on two arrays; > 80% reproducibility
MACROarray Reproducibility
Dilated Cardiomyopathy study, Horn et al. 2006
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� Using purified proteins printed on protein biochips
� 20 patient sera / 10 control sera from normal individuals
� 7 Disease control sera from patients with psoriasis, hand and foot
eczema or skin allergy
patient
sample 9
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sample 57
disease control
skin eczema
sample 5
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Diagnostic Protein Biochip: Alopecia areata
= Alopecia-associated proteins
Confirmation of MACROarray-identified antigens
7Protagen AG
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MACROarrays Used for Biomarker Discovery
• Systemic Lupus erythematodes (SLE mouse model)Gutjahr, C., ….. Cahill, DJ (2005) Mouse protein arrays from a TH1 cell cDNA library for antibody screening and serum profiling. Genomics 85, 285–296
• Alopecia areata (hair loss) Lüking A., …. Cahill, DJ (2005)
Profiling of alopecia areata autoantigens based on protein microarray technology. Mol. Cell Proteomics 4:1382–90
• Dilated Cardiomyopathy (DCM)Horn, S., …. Cahill, DJ (2006)Profiling humoral auto-immune repertoire of dilated cardiomyopathy (DCM) patients and development of a disease-associated protein chipProteomics 6: 605 - 613
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Protagen Patent Position
Patented UNIclone® platform technology
• Creation and use of protein expression libraries
• Use of recombinant products originating from expression
libraries in array formats
• Protagen has exclusive worldwide license
from Max-Planck-Society, Germany
Protagen’s patent position
• Unique and solid position for protein arrays
• Freedom-to-operate towards products &
cooperations in the Life Science industry
8Protagen AG
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A B C E F G HD
Capturing antibody
Detecting antibody
Secondary antibody
Label
Immobilized protein
Interacting protein Small compound
Protagen Protein Biochips
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Quantitative binding profiles of biotherapeutics
Antibody A Antibody B Receptor Fusion
Antigen Antigen Antigen
UNIchip® AV-400 UNIchip® AV-400 UNIchip® AV-400
Example: UNIchip® AV-400, Launch 2005
Patented design with antigen calibrator
9Protagen AG
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Antigen Discovery for Multiple Sclerosis
Systematic discovery of novel Multiple Sclerosis autoantigens
in a BMBF-supported project
• Identification of putative autoantigens using MACROarrays
• Screening patient cohorts using purified antigens on protein biochips
• Disease control: acute disseminated encephalomyelitis (ADEM), healthy controls
• Clinical partner:Dr. Sebastian SchimrigkNeurologische Klinik der Ruhr Universität Bochum, St. Josef HospitalArbeitsgruppe NeuroimmunologieTherapie und Studienzentrum für Multiple Sklerosis
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Discovery of Putative Multiple Sclerosis Antigens
MS patient serum on hEx1 MACROarray
10Protagen AG
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Development and production of protein biochips with
recombinant antigens for diagnostic multiplex assays, patented
• MACROarray processing
• Clone selection & identification
• High-throughput protein expression & purification
• Screening & bioinformatic analysis
• Manufacturing quantitative protein biochips
Unique Strength of Protagen
JB/26.11.2006 Slide: 20
Protagen AG
Otto-Hahn-Str. 1544227 DortmundGermany
T +49-231-9742 6300F +49-231-9742 [email protected]