Protein Microarrays and detection via MALDI-TOF: Project Overview - PowerPoint PPT Presentation

View by Category
About This Presentation
Title:

Protein Microarrays and detection via MALDI-TOF: Project Overview

Description:

Protein Microarrays and detection via MALDI-TOF: Project Overview Kyle Nordquist Advisor: Dr. Andrew Link Microarray Foundation DNA microarrays method of high ... – PowerPoint PPT presentation

Number of Views:120
Avg rating:3.0/5.0
Slides: 11
Provided by: nord168
Category:

less

Write a Comment
User Comments (0)
Transcript and Presenter's Notes

Title: Protein Microarrays and detection via MALDI-TOF: Project Overview


1
Protein Microarrays and detection via
MALDI-TOF Project Overview
  • Kyle Nordquist
  • Advisor Dr. Andrew Link

2
Microarray Foundation
  • DNA microarrays method of high-throughput gene
    analysis
  • DNA nucleotides with specific gene expression
    spotted
  • Chip probed fluorescently

3
Research Purposes
  • Different Interactions
  • DNA-DNA
  • DNA-small molecule
  • DNA-antibody

4
Protein Applications
  • Interactions at Protein level
  • Protein-Drug relationships
  • Crucial pharmaceutical tool

5
Complications
  • High-throughput purification --tedious
  • Proteins must remain in native conformation to be
    of any use
  • Binding affinities
  • Tagging (His/GST)

6
GST-purification
  • Yeast genome library obtained in 96-well format
  • Cells must be grown and lysed
  • Overexpressed proteins must be extracted
  • Contamination

7
Spotting
  • Vanderbilt Microarray Shared Resource Center
  • Numerous surface chemistries to be tested

8
Current Detection
  • Published literature using fluorescent detection
    data
  • Shows protein intensity
  • Fails to show
  • Size of proteins
  • Interaction affinity

9
MALDI-TOF MS
  • Matrix-Assisted Laser Desorption and Ionization
    Time-of-Flight Mass Spectrometry
  • Has not been used to quantify protein chip data

10
Possible Advantages of using a MALDI detection
system
  • Specific mass data in reference to binding
    interactions
  • Classify what molecules bound to what proteins
  • Easy to distinguish unknown proteins by weight
  • Less time for preparation
About PowerShow.com