METHOD FOR PREDICTION OF FUTURE CARDIOVASCULAR DISEASE RISK VIA ANALYSIS OF IgG GLYCOME

A method for prediction of future cardiovascular disease (CVD) in a human subject comprises performing an analysis process of N-glycans I, bound to immunoglobulin G <IgG>,

• where various symbols in I denote monomeric sugar units as below:
• ▪ N-acetylglucosamine
• ▾ fucose
• ● C mannose
• ∘ galactose
• ♦ sialic acid
• where letters a-d in I determine types of glycoside bond of said N-glycans I as below:
• a
• =
• β
• ⁢
• 〈
• 1
• -
• 4
• 〉
• b
• =
• α
• ⁢
• 〈
• 1
• -
• 6
• 〉
• c
• =
• α
• ⁢
• 〈
• 1
• -
• 3
• 〉
• d
• =
• β
• ⁢
• 〈
• 1
• -
• 2
• 〉
• where the analysis process comprises following steps:
• a) isolation of plasma from one or more blood samples that have been collected from the human subject under examination,
• b) isolation of IgG from blood plasma of the blood samples,
• c) release of said glycans from IgG,
• d) fluorescent derivatization with 2-aminobenzamide <2AB> and a reducing agent for reductive amination, optionally using a complex of picoline borane <BH3·NC5H4-2-CH3> or sodium cyanoborohydride <NaBH3CN>:
• e) quantitative analysis of thus derivatized glycans by ultraperformance liquid chromatography <UPLC>, wherein numerical values of relative areas under respective glycan peaks GP 9, GP 12, GP 19, and GP 20 within the corresponding UPLC chromatogram are obtained as the numerical values {GP9, GP12,GP 19, GP20}, wherein the said glycan peaks GP 9, GP 12, and GP 19 belong to structures given in the table below:
• Glycan peak composition
• Code
• Structure
• F(6)A2[3]G1 <100%>
• GP9
• A2G2 <91%> F(6)A2[3]BG1 <9%>
• GP12
• F(6)A2BG2S1 <100%>
• GP19
• wherein G20 represents a peak at a retention time calibrated to retention times of glucose oligomers labeled with 2-aminobenzamide,
• f) wherein numerical results of relative areas under the selected glycan peaks GP9, GP12, GP19, and GP20, that correspond to values {GP9, GP12, GP19, GP20}calculated in step e, are now included in a CVDR model for cardiovascular diseases risk <CVDR>, which is a function of the four glycan arguments:
• CVDR=CVDR(GP9,GP12,GP19,GP20)
• g) determination of the risk value expressed as the CVDR value.