Comparison of ultracentrifugation and gel filtration for the isolation of bovine lipoproteins |
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Authors: | R R Grummer C L Davis H M Hegarty |
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Affiliation: | (1) Department of Dairy Science, University of Illinois, 1207 W. Gregory Drive, 61801 Urbana, IL |
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Abstract: | Lipoproteins from the plasma of three nonlactating Holstein cows were isolated using either preparative ultracentrifugation
or gel filtration chromatography. Lipoprotein classes obtained by ultracentrifugation were very low density plus chylomicra,
<1.006 g/ml; low density, 1.007–1.039 g/ml; high density1, 1.040–1.063 g/ml; and high density, 1.064–1.22 g/ml. These lipoprotein classes were individually applied to an agarose gel
column to determine at what volume they eluted in comparison to lipoproteins that were separated after applying total bovine
lipoproteins to the column. Three major peaks corresponding to very low density lipoproteins plus chylomicra, low density,
and high density lipoproteins resulted after gel filtration of total lipoproteins. Very low density lipoproteins plus chylomicra,
obtained by ultracentrifugation, eluted as a single peak, as did low density and high density lipoproteins. However, high
density1 lipoproteins eluted as two peaks. The first peak eluted at the same volume as low density lipoproteins, and the second peak
eluted at a volume similar to that of the ascending slope of the high density lipoprotein peak. Results from disc polyacrylamide
gel electrophoresis, immunoelectrophoresis and double immunodiffusion of lipoprotein fractions, and SDS polyacrylamide gel
electrophoresis of their apoproteins, similarly indicated that the lipoproteins present in the 1.040–1.063 g/ml density interval
are a mixture of low and high density lipoproteins rather than a unique class of lipoproteins. |
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