1. Portable SPR instrument: Fromresearch to clinical applications Jean-François Masson Université de Montréal PhotonicsNorth 2014 – Parlons Affaires

2. Field of the innovation • Scientificequipment for the research, industrial applications and clinical diagnostics • Innovation: • Small and portable SPR instrument • Surface chemistry to analyzecrudebiofluids • Nanomaterial to amplifyresponsefrom instrument

3. Commercial SPR instrument – lab-based • Pros: High sensitivity, reliability and automation • Cons: High cost of acquisition, operation and maintenance, not possible to use biofluids

4. Commercial SPR instrument – Portable • Pros: portability, low acquisition cost • Cons: Poorer performance, maintenance, user-friendliness, low number of sensing channels (typically 1-2)

5. Integrated portable SPR instrument Features • Sensitive • 10-6 RIU resolution • Computer poweredwith USB – no external power cord • No RI matchingfluids • Simple injection system • 4-channels including a reference • Simplicity • Versatility • Performance • Portability • Customizable • Low maintenance • Lowcost • Lowweight (lessthan 1.3 kg) • Quiet operation

6. Industrial application – Salinitymeasurement Reproducibility < 1% and thechannel to channel variation < 10% • Calibration runwith 3 channels (A-C active), background subtractedwithchannel D • Triplicaterepeat of calibration point 2 at the end of the run R2 = 0.9998 % CV = 0.6 to 8% Repeatability: 0.4 % (6.02 ± 0,02) nm

7. Screening growth hormone-releasing peptides (GHRPs) Type B scavenger receptor CD36 (CD36: cluster of differenciation 36) • EP80317: anti-atherosclerotic property mediated by CD36 • interfering with the binding of oxLDL to the scavenger receptor expressed on macrophages

8. Therapeutic drug monitoring Methotrexate – chemotherapy agent • Drugswith a narrowtherapeuticwindow must becarefullycontrolled to ensurepatient’ssafety, propermetabolism and drugefficacy • Common analyticaltechniques are laboratory-based, time-consumingor expensive to run • The development of a smalland portable instrument with a simple and dedicated set of reagentswouldaddresssome of the current challenges in TDM

9. Quantification of MTX in clinicalsampleswith SPR instrument Collaboration with a local hospital to analyzeactualclinicalsamples • The samplesweredeidentified and analyzed in a blindassaywith FPIA (independantanalyst) and with LC-MS/MS and SPR (differentanalyst SZ) • FPIA analysisresultswereonlyknownafter final report from SPR and LC-MS/MS

10. Calibration fromdifferentoperators and locations Calibrations wererun in UdeMlaboratories and at local hospital • The system wasdeployedatHosp. Maisonneuve-Rosemont to evaluate the performance in a real environment • The SPR system is running on power supplied by the portable computer • Ease of portability 10 uM Day 1 CV = 4-8% 0 uM Day 2 CV = 6-16% 50 uM Day 2 CV = 9-23%

11. Acute lymphoblasticleukemia • Measuring the immune responseduringtreatment • Some patients respond to asparaginasetreatment by developingantibodies, leading to ineffective treatment and allergicreactions • Monitor antibody concentration by immolizing the drug (asparaginase) on the SPR sensor His-tag Asparaginase SPR sensor

12. Proteinbiomarkers • Prostate-specificantigen (PSA) Screening method: • Digital rectal exam: • Low cost • Unpleasant • Only screen a section of the prostate • Prostate specific antigen (PSA) test: • Protein secreted by the prostate cells • Level measure in a blood sample • Complementary to digital rectal exam foran accurate diagnostic ***Ratio of PSA + Ab-2 (1:1) used LD: 0,1 nM

13. Interfacingwithotherspectroscopic techniques • SPR-fluorescence Multi-channel SPR and fluorescence instrument: • SPR and fluorescence are orthogonal techniques thatcanbecombined in a small instrument Fluorescence Fluorescence SPR SPR

14. Proof-of-principlewith SPR-fluorescence bioassay • Fluorescent antibodyused: Cy3 Anti-Human-IgG • Usingmicrostructured gold film couldimprove the fluorescence emission Positive control (1000nM IgGimmobilized) SPR Fluorescence

15. Differentmarkets accessible • Research (ready to market) • Ligand screening • Bindingassays • Optical research / Surface-enhanced spectroscopies • Industrial applications (partiallyvalidated) • Refractive index measurement • Fermentation processes • Clinical diagnostics (in process ofvalidation) • Biomarkers • TDM

16. Acknowledgements Currentresearch group: Dr. Natalia Bukar Dr. Hélène Yockell-Lelièvre Dr. KristyMcKeating Dr. Marc Vidal Sandy Shuo Zhao * Julien B.-Turcot Maxime Couture Hugo-Pierre P-Richard Rita Faid Alexandra Aubé Hu Zhu Simon Forest Daniel Pelechacz * Félix Lussier Jérémie L.-Carbonneau Geneviève Granger Gabrielle L. Lajoie Corentin Geny * Alumni