Lipoprotein fingerprinting methods using metal ion chelate salts
AssigneeThe Texas A&M University System
InventorsRonald D. MacFarlane, Brian D. Hosken, Zachlyn N. Farwig, Irma L. Espinosa, Christine L. Myers, Steven L. Cockrill
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Abstract
Methods are disclosed which separate and identify lipoproteins in biological samples. An ultracentrifuge density gradient is used to separate lipoprotein fractions. The fractions are visualized, resulting in a lipoprotein profile. The fractions can be further analyzed by a wide array of laboratory and clinical methods. The lipoprotein profile can be used in clinical diagnoses and other medical applications.
Claims24
A method of obtaining the lipoprotein profile of a sample, the method comprising:
- contacting the sample, a density gradient forming agent, and a dye to produce an unseparated mixture, wherein the sample contains lipoproteins;
- subjecting the unseparated mixture to a centrifuge force field under conditions suitable to produce a separated mixture, wherein the separated mixture comprises multiple lipoprotein fractions;
- and
- visualizing the multiple lipoprotein fractions to obtain a lipoprotein profile.
A method of separating total lipoprotein in a serum sample from non-lipoprotein serum proteins, the method comprising:
- contacting a serum sample, a density gradient forming agent, and a dye to produce an unseparated mixture, wherein the serum sample contains lipoproteins and the unseparated mixture has a density of at least about 1.3 kg/m3;
- and
- subjecting the unseparated mixture to a centrifuge force field under conditions suitable to produce a separated mixture, wherein the separated mixture comprises sedimented non-lipoprotein serum proteins and non-sedimented lipoproteins.
A method of obtaining a differential density lipoprotein profile of a sample, the method comprising:
- contacting the sample, a density gradient forming agent, and a dye to produce a first unseparated mixture, wherein the sample contains lipoproteins;
- subjecting the first unseparated mixture to a centrifuge force field under conditions suitable to produce a first separated mixture, wherein the first separated mixture comprises a first set of multiple lipoprotein fractions;
- visualizing the first set of multiple lipoprotein fractions to obtain a first lipoprotein profile;
- contacting the sample and an antibody that binds to a lipoprotein fraction to form an antibody-lipoprotein complex;
- separating the antibody-lipoprotein complex from the sample to produce a reduced sample;
- contacting the reduced sample, a density gradient forming agent, and a dye to produce a second unseparated mixture;
- subjecting the second unseparated mixture to a centrifuge force field under conditions suitable to produce a second separated mixture, wherein the second separated mixture comprises a second set of multiple lipoprotein fractions;
- visualizing the second set of multiple lipoprotein fractions to obtain a second lipoprotein profile;
- and
- subtracting the second lipoprotein profile from the first lipoprotein profile to obtain a differential density lipoprotein profile.
Dependencies are inferred from the claim text.