HDL-ASSOCIATED PROTEIN EXTRACTION AND DETECTION

摘要

Provided herein are compositions, systems, and methods for extracting and detecting at least one HDL-associated protein (e.g., ApoA1) from a sample (e.g., plasma or serum sample). In certain embodiments, a strong organic acid and hydrophilic organic solvent are mixed with the sample; after centrifugation, the supernatant is transferred to a second container where it is mixed with a non-polar organic solvent; after centrifugation, the lower aqueous layer is transferred to a third container; and then at least a portion of the transferred aqueous layer is subjected to a detection assay such that at least one HDL-associated protein is detected.

主权项

1. A method of extracting and detecting at least one HDL-associated protein 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 greater than 50% 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) subjecting at least a portion of said bottom aqueous layer to a detection assay such that at least one HDL-associated protein is detected.