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Lipoprotein analysis by differential charged-particle mobility

A lipoprotein, charged technology, applied in the analysis of materials, biological material analysis, material electrochemical variables, etc., can solve the problems of wrong quantitative results, lipoprotein confusion, wrong qualitative results, etc.

Active Publication Date: 2010-07-07
QUEST DIAGNOSTICS INVESTMENTS INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, differential dye uptake may produce erroneous quantitative results
In addition, uneven color rendering can produce erroneous qualitative results because measured peaks can be distorted enough to confuse one class or subclass of lipoproteins with another class or subclass
Also, gradient gel electrophoresis can take many hours to complete

Method used

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  • Lipoprotein analysis by differential charged-particle mobility
  • Lipoprotein analysis by differential charged-particle mobility
  • Lipoprotein analysis by differential charged-particle mobility

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0107] Example 1 - Using D 2 Comparison of Purified Lipoproteins with O and Low Salt Solutions

[0108] Use a low-density salt solution (1.151 g / mL) (ie, "low-density salt sample") or D 2 Serum samples (25uL) were treated with O (200uL each). Samples were centrifuged at 223,000×G for 3.7 hours (low density salt samples) or 2 hours (D 2 O). After removing the top 100 uL after centrifugation, low density salt samples were dialyzed against ammonium acetate solution and diluted to 1:200 prior to differential charged particle mobility analysis. Before differential charged particle mobility analysis, D 2 O samples were directly diluted to 1:200 with ammonium acetate after centrifugation. The results of the differential charged particle mobility analysis are in figure 2 displayed in .

Embodiment 2

[0109] Example 2 - Effect of Purification on Apo A, Apo B and TC Recovery

[0110] To assess HDL (Apo A1) in the use of D 2 Whether the loss is preferred in the step of O, such as image 3 The three samples shown in (i.e., 749, 1043, 14: arbitrary and unique patient identification numbers) were subjected to lipoprotein separation using D 2 O along with the RGD / DS solution (7.5 / 2.5 mg / mL, respectively) solution to remove the albumin. Each sample was prepared in six replicates. The top 100 uL of each isolation was analyzed for Apo Al (HDL), Apo B (LDL, IDL, VLDL) and total cholesterol (TC) content. Plasma or serum apolipoproteins AI and B were measured by standard ELISA using a commercially available monoclonal capture antibody (Biodesign International, Saco, MN) and an anti-human goat polyclonal detection antibody in a noncompetitive sandwich immunoassay Apolipoproteins AI and B were purified and biotinylated (International Immunology Corp., Murrieta, CA). Concentrations w...

Embodiment 3

[0111] Example 3 - Effect of different RGDs on recovery of lipoprotein fractions

[0112] being paved in D 2 Serum samples were mixed with different amounts of RGD (10, 15, 20, 25 mg / mL) and incubated on ice for 15 minutes before topping the O cushion. After centrifugation at 223,000 x G for 120 minutes, the top 100 uL was removed and diluted 1:200 with ammonium acetate solution. The samples were then analyzed by differential charged particle mobility analysis. result in Figure 4 displayed in .

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Abstract

The invention provides apparatus and methods of preparation of lipoproteins from a biological sample, including HDL, LDL, Lp(a), IDL, and VLDL, for diagnostic purposes utilizing differential charged-particle mobility analysis methods. Further provided are methods for analyzing the size distribution of lipoproteins by differential charged-particle mobility, which lipoproteins are prepared by methods of the invention. Further provided are methods for assessing lipid-related health risk, cardiovascular condition, risk of cardiovascular disease, and responsiveness to a therapeutic intervention, which methods utilize lipoprotein size distributions determined by methods of the invention.

Description

field of invention [0001] The present invention relates generally to the field of particle size analysis and analysis of biological particles including lipoproteins for diagnostic purposes using ion mobility measurement devices and methods. The present invention also provides methods and apparatus for purifying and isolating biomolecules including, but not limited to, lipoproteins and biocomplexes containing lipoproteins. Background of the invention [0002] The following description is provided only to aid in the understanding of the present invention. None of the documents cited or information presented is admitted to be prior art to the present invention. [0003] Cardiovascular disease is the leading cause of death in the United States. In addition to patient demographics and current health, the most common and accepted methods for determining future heart disease risk include measuring serum concentration levels of cholesterol and lipoproteins. There are recommended ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/26
CPCG01N2800/32G01N33/92G01N33/5002
Inventor M·P·考菲尔德R·E·赖茨S·李G·K·李R·克劳斯P·J·布兰奇W·H·班纳E·考奈尔
Owner QUEST DIAGNOSTICS INVESTMENTS INC
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