Capillary Electrophoresis of Proteins and Peptides

345 Pages·2004·6.15 MB·English

by  Charles A. Lucy, Nicole E. Baryla (auth.), Mark A. Strege, Avinash L. Lagu (eds.)

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METHODS IN MOLECULAR BIOLOGY
Volume 276
Capillary
Electrophoresis
of Proteins
and Peptides
Edited by
Mark A. Strege
Avinash L. Lagu
Capillary Electrophoresis of Proteins and Peptides
M E T H O D S I N M O L E C U L A R B I O L O G Y™
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M E T H O D S I N M O L E C U L A R B I O L O G Y™
Capillary Electrophoresis
of Proteins and Peptides
Edited by
Mark A. Strege
Eli Lilly and Company, Indianapolis, IN
Avinash L. Lagu
Lilly Research Laboratories, Indianapolis, IN
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Preface
Throughout the more than 20 years that have followed the beginnings of
capillary electrophoresis (CE), its application to the analysis of proteins
and peptides has continued to be reliable, versatile, and productive. Over
time, CE has matured to become a superb complement to HLPC, and in
many cases has also evolved as an automated and quantitative replacement
for conventional slab gel electrophoresis methods such as SDS-PAGE and
isoelectric focusing.
Within Capillary Electrophoresis of Proteins and Peptides, we have
assembled contributions from researchers who are applying state-of-the-art
CE for protein and peptide analysis, including topics that we believe are of
great potential both in the present and for the future.
In comparison to traditional separation methods, CE represents a
miniaturized analysis technique (especially in its microchip-based format)
that is highly dependent upon the basic fundamentals of effective sample
recovery and high sensitivity detection. With these issues in mind, Chapters
1–4 describe recently developed approaches for both capillary coatings and
analyte detection via laser-induced fluorescence.
Since the discipline of biotechnology has established itself as a primary
platform for the application of CE to the analysis of proteins and peptides,
Chapters 5–7 demonstrate a variety of examples of the specific techniques
that have been applied for the development of biopharmaceuticals and their
commercialization. The methods covered here include also the analysis of
oligosaccharides from glycoproteins.
Studies of the association of proteins with other molecules can provide
insight into the very heart of biological processes. Therefore, a major focus
within both the pharmaceutical industry and academia is the utilization of
CE for the characterization of protein interactions with ligands, other
proteins, and large biopolymers. Chapters 8–11 describe in detail the most
recent approaches for performing affinity capillary electrophoresis for the
evaluation of protein binding, including the use of protein charge ladders.
CE and capillary isoelectric focusing have been providing rapid, high-
resolution separations of proteins. When combined with electrospray mass
spectrometry detection they constitute a powerful analysis system capable
v
vi Preface
of supporting complex studies such as those associated with proteomics.
Chapters 12–15 focus on the use of CE within this exciting field. The use of
CE in microfluidics format is also presented here.
The objective of Capillary Electrophoresis of Proteins and Peptides, by
its breadth, topicality, and forward focus, is to serve as a valuable guide for
researchers across many disciplines. We look forward with great
anticipation to the impact this collection will have, as researchers new to
the field are carried forward in their work by the experts’ step-by-step
guidance and notes provided within these chapters.
Mark A. Strege
Avinash L. Lagu
Contents
Preface .................................................................................................. v
Contributors ......................................................................................... ix
1 Surfactant-Based Methods for Prevention of Protein
Adsorption in Capillary Electrophoresis
Charles A. Lucy, Nicole E. Baryla, and Ken K.-C. Yeung............ 1
2 Capillary Coating for Protein Separation Based on Si-O
and Si-C Covalent Bond Formation for Capillary Electrophoresis
With Laser-Induced Fluorescence Detection
Hossein Ahmadzadeh, Norman J. Dovichi,
and Sergey Krylov ................................................................. 15
3 On-Column Labeling Reaction for Analysis of Protein Contents
of a Single Cell Using Capillary Electrophoresis
With Laser-Induced Fluorescence Detection
Hossein Ahmadzadeh and Sergey Krylov ................................. 29
4 Covalent and Noncovalent Labeling Schemes for Near-Infrared
Dyes in Capillary Electrophoresis Protein Applications
John Sowell, Jozef Salon, Lucjan Strekowski,
and Gabor Patonay ............................................................... 39
5 Capillary Electrophoresis in the Analysis and Monitoring
of Biotechnological Processes
Vadim Klyushnichenko ............................................................. 77
6 Capillary Electrophoresis of Proteins
in a Quality Control Environment
David L. Good, Stacey Cummins-Bitz, Raeann M. Fields,
and Brian K. Nunnally ......................................................... 121
7 Analysis of Neutral N-Linked Oligosaccharides
From Antibodies Using Free-Solution Capillary
Electrophoresis in Bare Fused-Silica Capillaries
Jeffrey S. Patrick, Brenda P. Rener, Gregory S. Clanton,
and Avinash L. Lagu ............................................................ 137
8 Affinity Capillary Electrophoresis to Examine
Receptor–Ligand Interactions
Maryam Azad, John Kaddis, Valerie Villareal, Lili Hernandez,
Catherine Silverio, and Frank A. Gomez .............................. 153
vii
viii Contents
9 Screening Major Binding Sites on Human Serum Albumin
by Affinity Capillary Electrophoresis
Hee Seung Kim, John Austin, and David S. Hage ................... 169
10 Using Charge Ladders and Capillary Electrophoresis
to Measure the Charge, Size,
and Electrostatic Interactions of Proteins
Upma Sharma and Jeffrey D. Carbeck ................................... 189
11 Frontal Analysis Continuous Capillary Electrophoresis
for Protein–Polyelectrolyte Binding Studies
Emek Seyrek, Toshiaki Hattori, and Paul L. Dubin ................. 217
12 Analysis of Proteins by CE, CIEF, and Microfluidic Devices
With Whole-Column-Imaging Detection
Jiaqi Wu, Xing-Zheng Wu, Tiemin Huang,
and Janusz Pawliszyn .......................................................... 229
13 Capillary Electrophoresis–Electrospray Ionization Mass
Spectrometry of Amino Acids, Peptides, and Proteins
Mehdi Moini ............................................................................ 253
14 Capillary Isoelectric Focusing–Mass Spectrometry
of Proteins and Protein Complexes
Suzana Martinovic´, Ljiljana Pasa-Tolic´,
and Richard D. Smith .......................................................... 291
15 Integrated System for Rapid Proteomics Analyses
Using Microfluidic Devices Coupled to
Nanoelectrospray Mass Spectrometry
Jianjun Li, Tammy-Lynn Tremblay, Jed Harrison,
and Pierre Thibault .............................................................. 305
Index ................................................................................................. 325
^
Contributors
HOSSEIN AHMADZADEH • Department of Chemistry, University of Minnesota,
Minneapolis, MN
JOHN AUSTIN • Department of Chemistry, University of Nebraska, Lincoln, NE
MARYAM AZAD • Department of Chemistry and Biochemistry, California
State University, Los Angeles, CA
NICOLE E. BARYLA • Eli Lilly Canada, Inc., Toronto, Ontario, Canada
STACEY CUMMINS-BITZ • Indianapolis Bulk Operations Technical Excellence
Centre, Eli Lilly and Company, Lilly Corporate Center, Indianapolis, IN
JEFFREY D. CARBECK • Department of Chemical Engineering, Princeton
University, Princeton, NJ
GREGORY S. CLANTON • Lilly Research Laboratories, Eli Lilly and Company,
Lilly Corporate Center, Indianapolis, IN
NORMAN J. DOVICHI • Department of Chemistry, University of Washington,
Seattle, WA
PAUL L. DUBIN • Department of Chemistry, Indiana University-Purdue
University at Indianapolis, Indianapolis, IN
RAEANN M. FIELDS • Indianapolis Bulk Operations Technical Excellence
Centre, Eli Lilly and Company, Lilly Corporate Center, Indianapolis, IN
FRANK A. GOMEZ • Department of Chemistry and Biochemistry, California
State University, Los Angeles, CA
DAVID L. GOOD • Indianapolis Bulk Operations Technical Excellence
Centre, Eli Lilly and Company, Lilly Corporate Center, Indianapolis, IN
DAVID S. HAGE • Department of Chemistry, University of Nebraska, Lincoln, NE
JED HARRISON • Department of Chemistry, University of Alberta, Edmonton,
Alberta, Canada
TOSHIAKI HATTORI • Research Center for Chemometrics, Toyohashi
University of Technology, Toyohashi, Japan
LILI HERNANDEZ • Department of Chemistry and Biochemistry, California
State University, Los Angeles, CA
TIEMIN HUANG • Department of Chemistry, University of Waterloo,
Waterloo, Ontario, Canada
JOHN KADDIS • Department of Chemistry and Biochemistry, California State
University, Los Angeles, CA
HEE SEUNG KIM • Department of Chemistry, University of Nebraska,
Lincoln, NE
ix