1 BioSystems International Lung Cancer Biomarker Test February 25th. 2011

2 BioSystems International oFounded in 2004 based on proteomics technology developed at Pfizer. oFacilities in Evry, France and Debrecen, Hungary. o35 Employees (23 in France, 15 in Hungary). oRecently merged with MicroBioChips, a French protein microarray company. Evry, France Debrecen, Hungary

3 Unique MAb Proteomics Biomarker Discovery Technology BSI has developed monoclonal antibody proteomics: a unique and superior technology for discovery of blood biomarkers: o Targets native epitopes: nascent mAB libraries ( mABs) cloned libraries (850 mABs) o Unbiased: No knowledge of disease mechanism or potential biomarkers assumed or needed. o High sensitivity: detects small changes in biomarker levels. o High-throughput: Biomarkers are confirmed by testing of hundreds of patient samples. o Efficient translation: Avoids major bottleneck of mass spectrometry- based biomarker discovery process: transition to clinical assay. o Diagnostic-grade MAbs against biomarkers are generated as part of the biomarker identification process; allows a rapid transition to clinical validation and IVD product development

4 Pipeline

5 Monoclonal antibody proteomics workflow Mass spectrometry based workflow for translation of protein biomarker discovery to clinical practice

6 BSI Biomarker Discovery Technology o Disease-state plasma samples are pooled and processed to remove the most abundant proteins and to “normalize” the concentration of remaining proteins o Mice are immunized with processed plasma samples o Resulting hybridomas are screened to select MAbs which discriminate disease state from control patient samples o Hybridomas secreting antibodies to candidate biomarkers are cloned. o Resulting MAbs used to identify biomarker o Libraries are searched for sandwich partners o Translation of discovery to diagnostic candidate, clinical testing

7 Targets of Antibodies oNative proteins which contain multiple epitopes oEpitope: a single antigenic site on a protein to which an antibody reacts o Linear or Discontinuous o Peptide o Carbohydrates oAntibodies detect PTRs Putative glycan epitope

8 More people die from lung cancer than any other type of cancer. Survival rate is ~16% but Why lung cancer / early diagnosis ? Average is 64.4%

9 Lung Cancer Discovery Workflow

10 Nascent hybridoma library: initial hits

11 Best candidates: cloned mABs

12 Cognate antigen identification Protein ID: immunoprecipitation  mass spectrometry  verification “redundancy” BM 4 BM1 BM2 BM3 BM2 BM3 BM5

13 Immunohistology: cancer tissue Negative control BM1 BM2 BM3 BM5

14 BM5: MAb-1 Recognizes Lung Cancer-Specific Epitope

15 Translation: Discovery Method to Sandwich ELISA Assays Disease specific isoforms? BM 1 BM 2 BM 3 BM 4BM 5

16 Lung Cancer Risk Indicator Panel oResults of 5 biomarkers were combined using support vector machine (SVM) algorithm. oLung cancer risk indicator is the composite linear function of individual biomarker concentrations. oGives improved sensitivity/specificity compared to any single biomarker.

17 Comparison of Individual Biomarkers to Panel

18 Risk Indicator Panel: performance BM 1-5

19 Febr CONFIDENTIAL Potential confounding effects Additional factors tested (capture assay) -Gender -Smoking habit -“Other” (non lung) cancer -Lung metastasis of “other” cancers SW ELISA: Cohort IV

20 Febr CONFIDENTIAL Combination with CYFRA : LC stages BM 1-4

21 Independent validation BM 1-5BM CYFRA

22 Conclusions, perspectives oBSI’s MAb proteomics technology identifies novel disease state biomarkers oThe new biomarkers combined with CYFRA have higher specificity and sensitivity for early LC than any previously reported biomarker panel oClinical use potential: o Diagnostic aid (current development focus) o Early diagnosis (partnering) o Management, monitoring (v2 panel is in the pipeline)

23 Acknowledgments Mariana Kuras Ph.D Istv á n Kurucz Ph.D. Nad è ge Tardieu M.S. William Hempel Ph.D Jozsef Lazar Ph.D. Yann Kiefert M.S. J á nos K á das Ph.D. Carole Malderez-Bloes M.S. Anne Jullien M.S. Andr á s Guttman Ph.D. Balazs Dezso M.D. Eszter Csanky M.D. Barry L. Karger Ph.D.. Mariana Kuras Ph.D. Biosystems International SAS, Evry, France Biosystems International Kft., Debrecen, Hungary Department of Pathology, University of Debrecen Department of Pulmonology, University of Debrecen Northeastern University Barnett Institute, USA Director of Research Biosystems Intl. SAS, France OSEO