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