Single-spin density-gradient ultracentrifugation vs gradient gel electrophoresis: two methods for detecting low-density-lipoprotein heterogeneity compared

1991 ◽  
Vol 37 (6) ◽  
pp. 853-858 ◽  
Author(s):  
Tom P J Dormans ◽  
Dorine W Swinkels ◽  
Jacqueline de Graaf ◽  
Jan C M Hendriks ◽  
Anton F H Stalenhoef ◽  
...  

Abstract Single-spin density-gradient ultracentrifugation (DUC) has proven to be a reproducible method for detection of low-density-lipoprotein (LDL) heterogeneity. Recently another method has been described for this: gradient gel electrophoresis (GGE) of serum, a method that might be more suitable for screening. To gain insight into the relationship of GGE to DUC and into their reproducibility, we determined LDL heterogeneity by DUC and GGE in 41 healthy individuals. In 90.2% (n = 37) of the subjects, the number of LDL subfractions found by both methods agreed. In addition, the density and the relative migration distance of the predominant LDL subfraction observed with the respective methods showed a strong correlation (Pearson correlation, r = 0.85, P less than 0.0001). Although it was not possible to compare for all aspects of LDL heterogeneity, these data suggest that GGE is a valid method of analysis for LDL heterogeneity. In screening programs, it may be necessary to store samples. Therefore, we studied in 24 sera the influence of storage at -80 degrees C for one, four, and 12 weeks on the LDL subfraction distribution detected by each method. LDL heterogeneity was maintained during storage under these conditions.

1990 ◽  
Vol 83 (1) ◽  
pp. 59-67 ◽  
Author(s):  
Bruce A. Griffin ◽  
Muriel J. Caslake ◽  
Brigitte Yip ◽  
Graeme W. Tait ◽  
Christopher J. Packard ◽  
...  

2003 ◽  
Vol 119 (3) ◽  
pp. 439-445 ◽  
Author(s):  
Shaina V. Hirany ◽  
Yusra Othman ◽  
Patricia Kutscher ◽  
David L. Rainwater ◽  
Ishwarlal Jialal ◽  
...  

Author(s):  
Hideko Tsukamoto ◽  
Izumi Takei ◽  
Keiko Ishii ◽  
Kiyoaki Watanabe

AbstractThe appearance of small, dense, low-density lipoprotein in serum has been demonstrated to be associated with increased risk of coronary artery disease. The molecular diameter of low-density lipoprotein is usually measured on the basis of mobility differences on polyacrylamide gel electrophoresis. However, since mobility assessed by this method is seriously affected by the increased levels of serum free fatty acids associated with hypertriglyceridemia, we used polyacrylamide gradient gel electrophoresis to eliminate the interference by fatty acids and devised a simple, precise method of polyacrylamide gradient gel electrophoresis to measure the diameter of small, dense, low-density lipoproteins in serum. We used apoferritin and thyroglobulin, which have a molecular diameter of 12.2 nm and 17.0 nm, respectively, and standard low-density lipoprotein particles having a diameter of 25.7 and 27.0 nm as calibrators, estimated by measurement of negative staining of electron microscopy. We also included apoferritin as an internal standard for polyacrylamide gradient gel electrophoresis. The only stain used was Coomassie brilliant blue, and it was used for lipoprotein staining. When we used low-density lipoprotein of 25.73 nm in diameter as a quality control specimen, the coefficient of variation of the size measurements obtained by our method was less than 1.2%. The new method markedly improved the laboratory procedure for measuring the diameter of low-density lipoproteins.


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