and chylomicrons, the polydispersity in size is much Gel filtration chromatographs of lipoproteins repremore severe. Gel filtration chromatography separates sent a superposition, or convolution, of the intrinsic solutes as a function of particle radius. There are sevpolydispersity of the solute and the dispersion due to eral practical advantages of gel filtration chromatogratransport phenomena. We describe a deconvolution phy as a method of analysis and preparation of lipoprotechnique for improving the resolution of gel filtration teins, including the rationality of the separation as a chromatographs applicable to lipoproteins and other function of particle radius, the nondisruptive nature of polydisperse solutes. A matrix of spreading functions, the matrix, and the use of ambient pressures and force characterizing the dispersive properties of the colfields, in contrast to those used in ultracentrifugation. umn, was determined by fitting chromatographic data A potential problem in gel filtration chromatography, from a series of monodisperse standards with the soluhowever, is that even monodisperse solutes (e.g., small tion to the transport equations and interpolating beions such as K / or Cl 0 ) do not elute at a single volume tween the fit parameters. A successive approximation (or time), but rather elute as a dispersion of elution scheme was used in which a test distribution was involumes about a modal value. In an ideal system, particrementally corrected by an amount proportional to cles would have a unique elution volume (or time) dithe error between the measured chromatograph and rectly related to the particle size (Fig. 1A). Practically, that derived from the test distribution. A nonlinear however, there is diffusion of particles, nonuniform relaxing function was used to constrain the correction flow around the beads making up the porous network, term such that the solution remained physically realizand the possibility of nonequilibrium between mobile able (i.e., nonnegative absorbance) as it evolved. Deand stationary phases. Furthermore, when a sample convolved chromatographs of lipoproteins provided contains more than one particle size, the spread peaks resolution of peaks that were obscured by spreading in the original data. The distribution of particle sizes due to each constituent may overlap, and the measured within each fraction was calculated and verified exabsorbance is the sum of these curves (Fig. 1B). This perimentally by further separating the contents of spreading of individual peaks can shift the position of fractions by gradient gel electrophoresis. Our techthe peaks or obscure them altogether. The spreading of nique, however, provided comparable resolution of the peaks depends on the properties of the filtration system peaks without the additional experimental procedure. such as the properties of the matrix, the flow rate, and