Conclusion And Future Perspectives

Lipoproteins, in addition to apolipoprotein identification and quantification, will continue to play an important role in the understanding of the pathogenesis of atherosclerosis for many years. Further development efforts will be directed towards the development of increasingly affordable, rapid, reliable, and automated technologies. CE with its various modes has several advantages when compared to the current procedures for lipoprotein and apolipoprotein analyses. These advantages are extreme analytical flexibility, using small quantities of low cost separation buffer, ability to use of a variety of detection modes, small sample size (nanoliter), high speed, efficiency, and reproducibility. It is also possible to install fully automated procedures. Furthermore, CE allows a rapid change from one buffer system to another: in other words, change from one analytical procedure to another.

In this chapter it becomes obvious that use of CE for lipoprotein analysis is still in its infancy. LDL, which is routinely only detected as one fraction, can be subdivided in up to six fractions by CITP that may yield new insights in the pathophysiology of the LDL-subfractions. It is possible that new areas in the study of atherogenesis may be achieved by the analysis of HDL subfractions. Since CE first separates then detects the lipoproteins, immuno-assay problems caused by antibodies are eliminated. In addition, since multiple apolipoproteins can be determined in one run, CE is a good alternative to procedures like gel electrophoresis or UC. In the future, with a multi-capillary instrument and with on-line sample pretreatment, it could well be competitive to automated special analyses. For broader acceptance of lipo-protein and apolipoprotein analysis by CE, three things are needed: 1) The development of chemical reagent kits, instruments, and software suitable for daily clinical laboratory use; 2) an increase of sensitivity and sample throughput; and 3) comparison and evaluation of various applications with current laboratory methods under daily laboratory conditions. In addition special attention needs to be paid to accuracy, reproducibility, speed, and susceptibility to disturbances (methods, capillaries, CE instruments). Other important criteria are ease of use in routine work situations, possibly auto mation, low cost per test, inexpensive and long-lasting reagents, widely accessible methodology, and clinically relevant reference ranges. Until now the number of reports evaluating these applications in comparison to the traditional procedures is low, however, this is likely to change in the near future.

Taken together, high-efficiency separations based on charge-to-mass ratio have been achieved through a combination of buffer modifiers and uncoated or permanently coated capillaries. Use of dynamically coated capillaries and separations based on the molecular weight have not yet been tested, but may further improve separation of the lipoproteins. CE offers more than simple improvement in quantitative and qualitative analyses of lipoprotein and apolipoproteins. It also offers the possibility of monitoring the reaction rates of apolipoproteins with different surfactants, lipoprotein degradation, lipo-protein metabolism, as well as therapeutic effects on lipoprotein classes. The use of CITP in the analysis of lipoproteins may facilitate the development of new clinical screening methods, possibly leading to new diagnostic markers or patterns, better prognostic evaluation, and new therapeutic modalities. Of course, as with all new developments, it is likely that some things may not turn out to be as important as expected. But I am also confident that many as yet unanticipated discoveries will be made as a result of the increased performance of this analytical technique. Nevertheless, the impact that these additional CE applications will have on diagnosis and treatment of atherosclerosis greatly depends on the success of transferring them from the academic research laboratories to the commercial marketplace.

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