Patent ID: 11940452
Assignee: CLEVELAND HEARTLAB, INC.
Field: Measurement (Instruments)
Classification: CPC G | IPC G

Claim 1:
2. A method comprising:
a) mixing, in a first container, a separating solution and a serum or plasma sample to generate a first mixed sample, wherein said separating solution comprises a strong organic acid and a hydrophilic organic solvent, and wherein said separating solution makes up 55%-93% of said first mixed sample;
b) exposing said first mixed sample to centrifugal force such that said first mixed sample separates into a pellet and supernatant;
c) transferring at least a portion of said supernatant to a second container such that it is separated from said pellet;
d) adding a non-polar organic solvent to said second container at a ratio of greater than 1:1 to generate a second mixed sample;
e) exposing said second mixed sample to centrifugal force such that said second mixed sample separates into an upper organic solvent layer and a bottom aqueous layer;
f) transferring at least a portion of said bottom aqueous layer to a third container such that it is separated from said upper organic solvent layer; and
g) detecting at least one HDL-associated protein in said bottom aqueous layer by subjecting at least a portion of said bottom aqueous layer to a detection method selected from the group consisting of, surface plasmon resonance, an activity assay, co-immunoprecipitation assay, Fluorescence Energy Transfer (FRET), bioluminescence energy transfer (BRET), an immunoassay, interferometry, Ellipsometry, and Quartz Crystal Microbalance, wherein said at least one HDL-associated protein is selected from Apolipoprotein A-I (APOAI), Apolipoprotein A-II (ApoA2), Alpha-1-antitrypsin (SERPINA1), Apolipoprotein C-III (APOC3), Apolipoprotein C-I (APOCI), Apolipoprotein A-IV (APOA4), Apolipoprotein C-II (APOC2), Angiotensinogen (AGT), Complement C3 (C3), Complement C4-B (C4B), Serum amyloid A-4 protein (SAA4), Transthyretin (TTR), Pigment epithelium-derived factor (SERPINF1), Apolipoprotein L1 (APOL1), Apolipoprotein F (APOF), Apolipoprotein M (APOM), Apolipoprotein Cl (APOC1), Fibrinogen alpha chain (FGA), Apolipoprotein E (APOE), Inter-alpha-trypsin inhibitor heavy chain H4 (ITIH4), Serum amyloid A-1 protein (SAA1), Alpha-2-HS-glycoprotein (AHSG), Complement C4-A (C4A), Retinol-binding protein 4 (RBP4), and Apolipoprotein D (APOD).