1. AgeLab Pharma GmbH, Hamburg, Germany *Dermatology Clinic, University Hospital, Eppendorf, Hamburg Amal K. Mukhopadhyay, Niels Grabe, Karsten Neuber *

2. Recombinant DNA Technology and Functional Genomics have given us an unprecedented possibility to diagnose, and sometimes even to treat many diseases which were so far categorised as of idiopathic origin.

3. In the post-human genome era, with nearly the entire human genome sequence being revealed, the proteomics technology has moved to a centre stage in the drug discovery processes.

4. Growth in Pharma companies has been driven by the technologies emerging from biotech industries Target identification Target validation Lead Identification Lead optimisation Preclinical developments Moving bottlenecks in the discovery value chain Adapted from Recap.com, L.E.K. analysis

5. "Life would be infinitely happier if we could only be born at the age of eighty and gradually approach eighteen." -- Mark Twain, Autobiography with Letters

7. Mega-Trend Aging

8. Predicted changes in the burden of disease from 1990 to 2020 Lower Respiratory infection 11Ischaemic heart diseaeses Diarrheal diseases22 Unipolar major depression Perintalassociated conditions33Road traffic accidents Unipolar major depression44Cerebrovascular diseases Ischaemic heart diseaeses55Chronic obstr. lung disease Cerebrovascular diseases66Lower respiratory tract inf. Tuberculosis77Tuberculosis Measles88War Road traffic accidents99Diarrheal diseases Congenital anomalies1010HIV 1990 2020

9. Non melanoma skin cancer stomach Bladder Colon Prostate Respiratory tract / Lung Rectum Stomach Uterus Breast Non-Hogkin Lymphoma Melanoma Kidney/Urinary Tract Stomach Ovary Non melanoma skin cancer Colon Rectum Melanoma Bladder Respiratory tract / Lungs Ten most common cancers in menTen most common cancers in women Common Types of Cancers ( Rheinland-Pfalz)

10. AgeLab: Organspecific Ageing with a focus on skin Section Young skin Section: Aged skin

11. Molecular analysis of ageing process Alterungsprozessen Young Old Sick Protein- profile Samples Difference

12. Kd 220 97 66 45 30 20,1 14,3 Increased Decreased 1 7 89 10 11 12 14 15 16 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 17 18 19 40 41 Y

13. Kd 220 97 66 45 30 20,1 14,3 Increased Decreased 1 7 8 9 10 11 12 14 15 16 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 17 18 19 40 41 O

15. Pooled Standard 2D gel co-separation IPGphor, Ettan DALT six Image acquisition Thyphoon 9400 Cy2/Cy3 co-detection & normalisation Cy2/Cy5 co-detection & normalisation Differential In-gel Analysis (DIA) DeCyder Software 3D protein profile Cy3Cy5Cy2 Mix samples Sample A control Sample B treated DIGE Cy-dye labelling 2D sample preparation 2D Clean-Up Kit 2D Quant Kit DIGE - experimental workflow

16. DeCyder differential in gel (DIA) analysis Result of full automatic spot detection: - 2153 spots detected - 270 spots decreased* - 170 spots increased* * at least 2-fold change of max spot volume; no spots excluded by filtering

17. DeCyder DIA analysis - Example 1 Spot 658 increased, volume ratio 3.93

18. DeCyder DIA analysis - Example 2 Spot 714 increased, volume ratio 6.02

19. DeCyder DIA analysis - Example 3 Spot 1685 increased, volume ratio 7.28

20. The ProteomeWorks „ Workflow“ IMAGING PDQUEST 2-D GELS OR BLOTS SAMPLE PREP ProteinLynx PepSeq PROTEIN DIGEST STATION MASS SPEC MALDI-TOF MS/MS ESI-MS SWISS-PROT TrEMBL,etc. HTML Links SPOT CUTTER

21. Marker-Proteins for Aging 6 years 34 years 78 years 39 years M 1 M 2 81 years

22. Regulatory Network of Cells Systems-Biology Bioinformatics based diagnostics Protein-Chip

23. Minimal-invasive Micro-biopsy for sample collection Suitable for epidermal diseases - Dermatitis - Eczema - Epidermal skincancer - Skin cancer precursor cell abnormalities. New Diagnostic procedure

24. - Chip is coated with specific antibodies against identified markers - AgeLab has identified more than 50 different potential marker proteins. –Apoptosis regulatory proteins (13) –Cell cycle regulatory proteins (9) –Nitric oxide (NO) signaltransduction regulatory Proteins (5) –Intermediate Filament-Proteins (13) –Chaperones (7) –Tumor Suppressors (8) Chip-Substrat AgeLabs diagnostic Protein-Chip

25. Tissue sample was labelled with fluorescent dye and after incubation, the chip was scanned in a fluorescence scanner. Protein Chip Development

26. Early detection of epidermal skin cancer Currently punch-biopsy followed by histology is the only procedure for diagnosis of actinic keratoses, a pre- cancerous stage. The procedure is painful, leaves scar and the outcome is unfortunately not absolutely dependable. With the help of AgeLab's procedure which is free from any scar development, a diagnostic procedure is being offered for the first time which is patients-friendly and dependable results can be expected. Early diagnosis of pre-cancerous stage in skin becomes possible with the help of the protein chip based micro- diagnostic procedure developed by AgeLab.

27. Euro-India Biotech-Conference, Hamburg, September, 2004 Contact email: am@agelabpharma.de

28. Expression of Thanks University of Hamburg Prof. F.A. Leidenberger AgeLab GmbH: Dr. Matthias Koenig Mr. Björn. Ehlers, Ms. Monika Kistler Special thanks are due to Amersham Biotech, Freiburg and Bio Agency, Hamburg