Two-Dimensional Difference in Gel Electrophoresis for Biomarker Discovery - Proteomic and Metabolomic Approaches to Biomarker Discovery - page 185

CHAPTER
12

Two-Dimensional Difference in Gel
Electrophor esis for Biomarke r Discovery

Haleem J. Issaq, Timothy D. Veenstra
Laboratory of Proteomics and Analytical Technologies, Advanced Technology Program and
Frederick National Laboratory for Cencer Research, Frederick, MD, USA

OUTLINE

Introduction 191 Application of 2D-DIGE to Biomarker

Discovery 194

Gel Electrophoresis: Historical Perspective 192

Conclusions 195

Two-Dimensional Differential In-Gel
Electrophoresis

192 Acknowledgment 195

Strengths and Weaknesses of 2D-PAGE
and 2D-DIGE

References 195

193

INTRODUCTION

analytical methods for the separation of proteins
are two-dimensional sodium dodecyl sulfate
polyacrylamide gel electrophoresis (2D-SDS-
PAGE) and two-dimensional differential in-gel
electrophoresis (2D-DIGE), both of which can
separate hundreds of proteins in a single experi-
ment. Advances in electrophoresis, such as DIGE
and the introduction of preformed pH gradient
immobilized gel strips with various isoelectric
ranges and mass spectrometry (MS) technolo-
gies, have enabled the detection of separated
proteins at much greater speed and sensitivity

Over the last two decades, there has been an
increase in the efforts to develop technologies
capable of separating and quantifying large
numbers of proteins expressed within a cell
system (i.e., the proteome) with the hope of iden-
tifying proteins that can be used as disease
markers. The complexity of the proteome has
made developing methods for ef

cient separa-
tion and sensitive detection of proteins a critical
component of this effort. The most widely used