Tina quant lipoprotein a gen 2

To validate the proteomics data, we selected altered plasma proteins and cholesterol fractions for biochemical assays in individual samples from patients and healthy volunteers. For analysis, the COBAS c311 automated system was used. Cholesterol fractions and TRIGLycerides were estimated, per the manufacturer’s protocol, using enzymatic and colorimetric methods (CHOL HICo Gen.2, HDL-C Gen.3 and TRIGL, Roche, USA). Plasma lipoprotein, apolipoprotein A-1 and apolipoprotein B were quantified, per the manufacturer’s protocols, using immunoturbidimetric methods (Tina-quant Lipoprotein (a) Gen.2, Tina-quant Apoliprotein A-1 ver.2 and Tina-quant Apoliprotein B ver.2, Roche, USA).
PON-1 and haptoglobin plasma levels were quantified by ELISA. PON-1 was measured with a human total PON1 DuoSet^® IC (DYC5816-2, R&D Systems, USA) and haptoglobin with a human haptoglobin immunoassay Quantikine^® ELISA (DHAPG0, R&D Systems, USA), following the manufacturer’s instructions.

Iodixanol was supplied as 60% w/v solution (OptiPrepTM) by Axis-Shield PLC (Dundee, UK). OptiSealTM tubes (3.2 ml) were supplied by Beckman Coulter (Krefeld, Germany). Triacylglycerol, Cholesterol, LDL-Cholesterol, HDL-Cholesterol, apoB, apoA1 and lipoprotein(a) assays, calibrators and serum lipid controls were purchased from Roche (Mannheim, Germany). For lipoprotein(a), the measuring range was 7–240 nmol/L (Tina-quant^® Lipoprotein (a) Gen. 2, Roche). All lipid measurements were performed on the Cobas C501 from Roche. Inter- and intra-assay coefficients of variation for lipoprotein(a) are shown in Table S5. Non-HDL-C was calculated as the difference between TC and HDL-C. Glucose was measured on the Siemens ADVIA 2400. HbA1c was determined with the Tosoh G8 HPLC Analyzer. Hs-CRP was measured on the BN II Siemens nephelometric analyser. For fibrinogen, the ACL TOP (Instrumentation Laboratory) was used.

Fasting venous blood samples were collected for biochemical and genetic testing. Serum total cholesterol, triglyceride, high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol concentrations (LDL-C) were measured using an enzymatic colorimetry Roche Cobas c702 analyzer with inter-assay CVs <1.5%. Plasma lipoprotein(a) level [Lp(a)] was measured using molar concentration by particle-enhanced turbidimetric immunoassay with Tina-quant Lipoprotein(a) Gen.2 (Latex) Roche. Serum insulin and C-peptide were measured using an Abbott Alinity analyzer. The homeostasis model assessment index of insulin resistance (HOMA-IR) was calculated from the fasting plasma glucose and insulin concentrations [16 (link)]. Adipokines (leptin, total adiponectin, chemerin) were measured using Quantikine^TM ELISA kits (R&D Systems, Minneapolis USA) with intra and inter-assay CV ≤ 7%. Genomic DNA was extracted from peripheral blood leukocytes using desalting method.