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

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!!