Yee tchao phat l. tran David you Jeong-yeol yoon

1. E-beam Size-Dependent Self-Assembly Protein Array Yee tchaophat l. tranDavid you Jeong-yeol yoon

2. Protein Arrays • Detect protein-protein interactions • Identify substrates of protein kinases • Most commonly used for antibody detection • Used in biomedical applications to determine presence/ amount of proteins in biological samples

3. Protein Arrays • More diverse than DNA/RNA • Cellular physiology • Disease and cancer at the proteomic level

4. E-beam Lithography • Scanning a beam of electrons in an organized fashion across a surface covered with a film • Exposed/non-exposed regions can be selectively removed • E-beam comes from the Scanning Electron Microscope (SEM)

5. Concept Conjugation of antibodies: + + Beads-COOH IgG BSA ++++++++++++++++++ Silicon Wafer 180nm 50nm 90nm Generation of micro and nano-wells: PMMA + + + + + + + + + + + + + + + + + + + + ++ + + + + + + + + + + + ++ Silicon Wafer PMMA ++++++++++++++++++++ Silicon Wafer

6. Computer-Guided Vibrator

7. ResultsCharacterization of protein nanoarray

8. ResultsSize-dependent self-assembly of protein nanoarray 5 μm

9. ResultsHigh density nanoarray

10. ResultsFluorescence detection of mouse IgG Green emission indicates target binding

11. Limitations www.the-scientist.com/article/display/11822 • Proteins and antibodies are sensitive to environment • Low density nanoarray • Multiple molecules at defined positions when developing a multi-component assay • Biomarkers present at low concentrations

12. Conclusion • Fabricated high-density, multi-component nanoarray through E-beam lithography and size-dependent self-assembly • Detected mouse IgG in array format through fluorescence properties

13. Future Directions • Detect Octamer-4 and other proteins under a multi-component nanoarray from real culture media • Reduce array scale to detect single molecule Lehnert et al. J. Cell Science (2004)

14. Thank You!!