Fluorometric sedimentation equilibrium for lipoprotein sub-class analysis.

Abstract

Fluorometric density gradient ultracentrifugation is used to measure the lipoprotein density profile for cardiovascular disease risk assessment. The work presented establishes the effectiveness of using a single-spin separation as both an analytical tool and a preparative tool, while yielding valuable density information. This research expands on the analytical power of density gradient ultracentrifugation (DGU) by combining novel ethylenediaminetetraacetic acid (EDTA) gradient solutions, a fluorescent probe for analysis, and modern statistical methods for classification of heart disease risk. Sub-classes of lipoproteins are analyzed based on their density from the fluorescent lipoprotein density profile. The application of linear discriminant analysis (LDA) and sliced average variance estimation (SAVE) to the fluorometric DGU data yields a powerful classification tool. This method is capable of determining differences between control and cardiovascular disease patients that do not exhibit the traditional risk factors. The combination of these methods has great potential to serve as analytical tools for researchers in understanding the mechanisms of disease development and as a diagnostic tool for clinicians.