Rapid Strides In The Field Of Capillary Electrophoresis
High performance liquid chromatography (HPLC) has undergone a tremendous development
in the past two decades.This growth has been possible because of the advent of high-performance packing materials, suitable instrumentation, and a thorough theoretical understanding of separations.
We are seeing a similar development in the field of capillary electrophoresis(CE).The development in CE of capillary pretreatment procedures and permanent coatings is similar to developments in chromatography decades ago.Research in CE""in its present form with fused-silica columns and on-column detection""is more than a decade old (Jorgenson and Lukacs, 1981, 1983), and commercial CE instruments have been available since 1988.
The field is moving rapidly, illustrated by the fact that already six annual symposia on CE have been held and a number of comprehensive textbooks (see, for example, the textbooks edited by Camillieri, 1993; Landers, 1994; and Guzman, 1993)have been published recently. The reviews by Karger (1993) and Landers et al. (1993) capture the state of the art of CE. A sampling of the type of proteinrelated applications for which CE has been utilized in analytical biochemistryand clinical/diagnostic assays is given below.
Purity Assays
" QC of a manufacturing process
" Quantitation of contaminants or excipients
" Screening of samples prior to protein sequencing
Structural Studies
" Peptide mapping
" Two-dimensional methods, i.e., HPLC-CE, IEC-CE
" Microheterogeneity of complex proteins e.g., glycoproteins,monoclonal antibodies, histones,transferrins
" Oligomerization
Binding Studies
" Calcium, zinc-binding proteins
" Antigen-antibody immune complexes
" Drug""protein complexes
" Protein""DNA complexes
Process Analysis
" Quantitation of the final product
" Enzymatic digestion monitoring
" Derivatization (labeling) monitoring
" Deamidation, disulfide bridge reduction
" Conformational changes
Stability Studies
" pH, environment, temperature effects
Mobility Measurements
" (Semi-)empirical models for prediction of separation
Clinical/Diagnostic
" Serum protein assays
" Hemoglobin variants; globin chains
" Neuropeptide isolation from brain tissue
" Apolipoproteins
" Cytokines, transferrins, metallothioneins
" Proteins in single cells
" Isoenzymes
As in HPLC, various separation modes are applicable to the analysis ofproteins and peptides:
capillary zone electrophoresis(CZE), micellarelectrokineticcapillary chromatography (MECC), capillary isoelectric focusing(
CIEF), capillary gel electrophoresis (CGE), and displacement electrophoresisor isotachophoresis (ITP).
While in CE, the separation of small peptides(in the CZE mode) often is relatively straightforward and well understood, itappears that no single strategy is applicable for large peptides and proteins.As might be expected, this is largely due to the wide diversity and complexityassociated with these biomolecules.
Thus, different strategies often work fordifferent protein separation problems, hence requiring different CE separationmodes.
Reference-
https://www.beckmancoulter.com/wsrportal/bibliography?docname=727484-CEPrimer5.pdfby:Scott eric
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