1. E-beam Size-Dependent Self- Assembly Protein Array

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

6. Computer-Guided Vibrator

7. Results Characterization of protein nanoarray

8. Results Size-dependent self-assembly of protein nanoarray 5 μm

9. Results High density nanoarray

10. Results Fluorescence detection of mouse IgG Green emission indicates target binding

11. Limitations  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 www.the-scientist.com/article/display/11822

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!!