1. Protein Separation Cathy Castellon BME 273 Advisor: Dr. Haselton
Graduate Advisor: Greg Stone
9/18/2018

2. Proteomics: Current Technology
The need and desire to understand total protein expression
Relies on the microchemical characterization of peptides separated by 2D protein electrophoresis.

3. How does 2DE work? Separates proteins by isoelectric point (pI)
And second by size (molecular weight) using sodium didecyl sulfate polyacrylamide gel electorphoresis (SDS-PAGE)

4. Application of 2DE
To compare the expression of protein profiles from an arbitrary reference state of a cell, tissue, or organism, to the profile of an non-standard condition
Example: Exposure of rat kidney to lead alters:
76 proteins in cortex
13 proteins in medulla
Separate complex protein mixtures into their individual polypeptide components

5. Improvements Mass Spectrometry with isotope labeling
Using isotope-coded affinity tags
Molecular Scanner
takes 2DE gels and combines protease digestion and electroblotting to a membrane in a single step

6. Our Thoughts Microfluid Technique Uniformly hydrophobic slide
Create flow channel (lithography)
Create inlet and outlet points
Load fluorescently labeled protein solution into one end
Pump buffer solution through the channel
Fluoremeter will detect separation

7. Current Work Produce hydrophobic/phillic gradient slides
Use Si-lane glass slides
Measure Contact Angles
This will enable us to determine the hydrophobicity of a particular protein

8. Slide III (1-25-2002) Hydrophobic gradient3 4 6 5 1b 2b 3b 4b 5b 6b

9. Current Plan Microfluid Chamber shallow and relatively wide
increase surface area interaction
ridges that mimic chamber below

10. Future Work
Come up with a simple experiment to pretest our assumptions.
Create gradient
Create microchannel
mix protein solution back and forth
fluorescent scanner