e-beam size-dependent self- assembly protein array
TRANSCRIPT
E-beam Size-Dependent Self-Assembly Protein Array
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
Protein Arrays More diverse than DNA/RNA Cellular physiology Disease and cancer at the proteomic level
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)
Concept
5
+
PMMA
Silicon Wafer+ + + + + + + + + + + + + + + + + + + + ++ + + + + + + + + + + + ++
+
Beads-COOH
IgG BSA
Silicon Wafer ++++++++++++++++++
PMMA
Silicon Wafer ++++++++++++++++++++
180nm
90nm50nm
Computer-Guided Vibrator
ResultsCharacterization of protein nanoarray
ResultsSize-dependent self-assembly of protein nanoarray
5 μm
ResultsHigh density nanoarray
ResultsFluorescence detection of mouse IgG
Green emission indicates target binding
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
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
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)
Thank You!!