Protagen AG

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Protagen AG
Successful Strategy for Systematic Identification of Novel Biomarkers
Technologieforum Diagnostik – Frankfurt, 27./28. Nov. 2006
Dr. Jens Beator
Director Protein Biochips
Jens.beator@protagen.de
Protagen AG
Otto-Hahn-Str. 15
44227 Dortmund
+49-231-9742 6300
Fact Sheet
Protagen AG
Established:
1997
Legal form:
Corporation on stocks „Aktiengesellschaft“
Location:
Otto-Hahn-Str. 15
44227 Dortmund
Germany
Employees:
34
Industrial partners:
- Bruker Daltonik GmbH, Bremen
- Vivascience AG, Hannover
- Serva Electrophoresis GmbH, Heidelberg
JB/26.11.2006 Slide: 2
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Protagen AG
Products & Services - Overview
JB/26.11.2006 Slide: 3
Business Strategy
Horizon 3
Value / Revenues
Diagnostics
and
Therapeutics
Horizon 2
Protein Biochip
Production
Horizon 1
FeeFee-forfor-service
and BioBio-IT
2000
2004
2008
2010
Time
JB/26.11.2006 Slide: 4
Protagen AG
Biomarkers in Autoimmune Diseases
> 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
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?
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Protagen AG
The Unique Resource for Biomarker Discovery
hEx1 Protein Expression Library
• Derived from human fetal brain
tissue with largest diversity of expressed genes
193.500 human protein expression clones with His-tags
bacterial clones grown on PVDF filters
inducible protein expression
• 37.200 protein expressing clones
react 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
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Unique Strength of the Technology
MACROarrays:
the only product and the largest human protein diversity for
reproducible, unbiased screening
• hEx1 Protein Expression Library (other libraries available)
• Manufactured using robotic systems
Full traceability & identification of clones
Reproducible screening product with 16,000 different expression clones
• Low sample consumption (0.5 ml/sample)
• Large amount of antigen available for antibody binding
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Protagen AG
Identification of Biomarkers - Strategy
Patient and
control samples
Screening of 37,000
human expression clones
Bioinformatical Analysis
Factor19
Factor20
Factor21
Factor
20
Factor22
18
Factor23
16
Factor24
14
Factor1
12
Patient
Sample
Control
Sample
Factor2
10
Factor3
Factor4
8
Factor5
Factor6
6
4
Factor7
2
Factor8
0
Protein 1 Protein 2 Protein 3 Protein 4 Protein 5 Protein 5 Protein 6 Protein 7 Protein 8 Protein 9
Putative auto-antigens
Indication-specific
multiplex biochip
Generation of a putative
disease specific protein
subset
Factor9
Factor10
Factor11
Bioinformatical analysis
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MACROarray Serum Screening: Alopecia areata
AP
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.
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Protagen AG
MACROarray Reproducibility
2
1
1 and 2 show the results of two separate screens of the same
patient, screened separately on two arrays; > 80% reproducibility
Dilated Cardiomyopathy study, Horn et al. 2006
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Diagnostic Protein Biochip: Alopecia areata
Confirmation of MACROarray-identified antigens
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
control
sample 57
10
7
4
1
11
8
5
2
12
9
6
3
disease control
skin eczema
sample 5
10
7
4
1
10
7
4
1
11
8
5
2
11
8
5
2
12
9
6
3
12
9
6
3
= Alopecia-associated proteins
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Protagen AG
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 diseaseassociated protein chip
Proteomics 6: 605 - 613
JB/26.11.2006 Slide: 13
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
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Protagen AG
Protagen Protein Biochips
A
B
C
E
D
Capturing antibody
Secondary antibody
Detecting antibody
Label
F
H
G
Immobilized protein
Interacting protein
Small compound
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Example: UNIchip® AV-400, Launch 2005
Quantitative binding profiles of biotherapeutics
Antibody A
Antibody B
Antigen
UNIchip® AV-400
Antigen
UNIchip® AV-400
Receptor Fusion
Antigen
UNIchip® AV-400
Patented design with antigen calibrator
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Protagen AG
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 Schimrigk
Neurologische Klinik der Ruhr Universität Bochum, St. Josef Hospital
Arbeitsgruppe Neuroimmunologie
Therapie und Studienzentrum für Multiple Sklerosis
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Discovery of Putative Multiple Sclerosis Antigens
MS patient serum on hEx1 MACROarray
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Protagen AG
Unique Strength of Protagen
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
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Protagen AG
Otto-Hahn-Str. 15
44227 Dortmund
Germany
T +49-231-9742 6300
F +49-231-9742 6301
www.protagen.de
info@protagen.de
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