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Deconvolution of Gel Filtration Chromatographs of Human Plasma Lipoproteins
Journal article   Peer reviewed

Deconvolution of Gel Filtration Chromatographs of Human Plasma Lipoproteins

Kenneth A Barbee, Julie A Morrow and Stephen C Meredith
Analytical biochemistry, v 231(2)
Nov 1995
PMID: 8594977

Abstract

Gel filtration chromatographs of lipoproteins represent a superposition, or convolution, of the intrinsic polydispersity of the solute and the dispersion due to transport phenomena. We describe a deconvolution technique for improving the resolution of gel filtration chromatographs applicable to lipoproteins and other polydisperse solutes. A matrix of spreading functions, characterizing the dispersive properties of the column, was determined by fitting chromatographic data from a series of monodisperse standards with the solution to the transport equations and interpolating between the fit parameters. A successive approximation scheme was used in which a test distribution was incrementally corrected by an amount proportional to the error between the measured chromatograph and that derived from the test distribution. A nonlinear relaxing function was used to constrain the correction term such that the solution remained physically realizable (i.e., nonnegative absorbance) as it evolved. Deconvolved chromatographs of lipoproteins provided resolution of peaks that were obscured by spreading in the original data. The distribution of particle sizes within each fraction was calculated and verified experimentally by further separating the contents of fractions by gradient gel electrophoresis. Our technique, however, provided comparable resolution of the peaks without the additional experimental procedure.

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Web of Science research areas
Biochemical Research Methods
Biochemistry & Molecular Biology
Chemistry, Analytical
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