Apolipoprotein fingerprinting technique and methods related thereto

A method for determining the concentration of specific apolipoproteins in a biological sample, the sample being obtained from a patient and the sample including plasma, serum, and lipoprotein fractions, the method comprising the steps of;

• a) adding a specific volume of an internal standard to the sample;
• b) applying the sample to a surface-enhanced, Protein G-coated, antibody-bound chip and removing unbound sample components;
• c) analyzing the sample by mass spectrometry;
• and
• d) determining the concentration of the apolipoproteins using values of internal standards.

A method for use in assessing risk factors, disease state, drug treatment and response in the field of pharmacogenetics, comprising the steps of:

• a) collecting a first biological specimen from a human being;
• b) generating a first apolipoprotein fingerprint for the biological specimen, said apolipoprotein fingerprint comprising data on presence and absence of a plurality of targeted apolipoproteins, on structural features and differences between isoforms of said plurality of targeted apolipoproteins, and on concentration of said plurality of targeted apolipoproteins;
• c) comparing said first apolipoprotein fingerprint data to reported risk factor data to assess risk factors, or to reported disease state data to assess disease state or status;
• d) collecting a second biological specimen from the same human being;
• e) generating a second apolipoprotein fingerprint;
• and
• f) comparing said second apolipoprotein fingerprint data with said first apolipoprotein fingerprint data to assess drug treatment and response, where said second biological specimen is collected after a drug treatment.

A method for obtaining a profile of a plurality of selected apolipoprotein biomarkers in a biological sample from a subject, comprising the steps of:

• a) providing the biological sample from a subject;
• b) applying the biological sample to the surface of a capture reagent that specifically captures the plurality of selected apolipoprotein biomarkers, allowing the plurality of selected apolipoprotein biomarkers to bind to the capture reagent, and removing unbound sample components;
• c) analyzing the captured plurality of selected apolipoprotein biomarkers by laser desorption/ionization mass spectrometry;
• and
• d) preparing the profile using the analysis results,
• wherein the plurality of selected apolipoprotein biomarkers is selected from the group consisting of Apo CI, Apo CII, Apo CIII, Apo CIV, Apo A1, Apo A11, Apo A1V, Apo AV, Apo B48, Apo B100, Apo D, Apo E, Apo F, Apo G, Apo H, Apo J, Apo L, Apo M, isoforms thereof, and combinations thereof.

An assay for individualization, differentiation and relative quantification of a mixture comprising apolipoprotein isoforms, comprising the steps of:

• a) preparing a surface with an antibody reactive molecule;
• b) applying a plurality of apolipoprotein antibodies to said surface to bind with said antibody reactive molecule;
• c) applying a mixture of apolipoprotein isoforms to bind with said plurality of apolipoprotein antibodies, wherein said mixture of apolipoprotein isoforms may be non-purified and said plurality of apolipoprotein antibodies may be any molecular fragment adapted to bind as an apolipoprotein antibody;
• d) generating a phenotypic isoform profile of said mixture of apolipoprotein isoforms;
• and
• e) utilizing said phenotypic isoform profile for identification of status of a patient relative to a disease or condition, for identification of a compromised physiological process, or for definition of one or more disease specific apolipoprotein biomarkers or profiles.