Plasma Biomarkers in Fasting Mice

Fasting (5 h) blood samples were taken from the tail vein to prepare EDTA plasma. Blood glucose was determined during blood sampling using a hand-held glucometer (Freestyle Freedom Light, Abbott Laboratories, Lake Bluff, IL, United States). Plasma insulin, leptin, adiponectin, cholesterol, triacylglycerols, ALT, CK-18M30 were determined as described (16 (link)). ELISA assays were used to measure plasma serum amyloid A (SAA; KMA0021 Thermo Fisher Scientific, Waltman, United States), MCP-1/CCL2 (DY479 R&D, Minneapolis, United States), MIF (DY1978 R&D, Fetuin A (DY1563 R&D) and LPS binding protein (LBP; HK205-02 Hycult Biotech, Uden, Netherlands) according to manufacturer’s protocol.
Lipoprotein profiles were analyzed by first separating lipoproteins into fractions using fast protein liquid chromatography (FPLC) with an AKTA apparatus (Pharmacia, Roosendaal, Netherlands), as previously described (17 (link), 18 (link)). Subsequently, total cholesterol and triacylglycerols were measured in the fractions collected for profiling with enzymatic assays (Roche diagnostics, Basel, CHF) (19 (link)–21 (link)).

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Gart E., Salic K., Morrison M.C., Giera M., Attema J., de Ruiter C., Caspers M., Schuren F., Bobeldijk-Pastorova I., Heer M., Qin Y, & Kleemann R. (2022). The Human Milk Oligosaccharide 2′-Fucosyllactose Alleviates Liver Steatosis, ER Stress and Insulin Resistance by Reducing Hepatic Diacylglycerols and Improved Gut Permeability in Obese Ldlr-/-.Leiden Mice. Frontiers in Nutrition, 9, 904740.

Publication 2022

Freestyle Freedom Light KMA0021 MCP-1/CCL2 HK205-02

Adiponectin Assays Blood Blood glucose Ccl2 Cholesterol Diagnostics Edta Elisa Enzymatic assays Fetuin Held Insulin Leptin Light Lipoproteins Liquid chromatography Lps binding protein Plasma Protein Tail Triacylglycerols Vein

Corresponding Organization : Netherlands Organisation for Applied Scientific Research

Other organizations : Netherlands Metabolomics Centre, Leiden University Medical Center, BASF (Germany)

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Variable analysis

independent variables

Fasting (5 h)
Blood samples taken from the tail vein

dependent variables

Blood glucose
Plasma insulin
Plasma leptin
Plasma adiponectin
Plasma cholesterol
Plasma triacylglycerols
Plasma ALT
Plasma CK-18M30
Plasma serum amyloid A (SAA)
Plasma MCP-1/CCL2
Plasma MIF
Plasma Fetuin A
Plasma LPS binding protein (LBP)
Lipoprotein profiles (total cholesterol and triacylglycerols)

control variables

EDTA plasma preparation
Hand-held glucometer (Freestyle Freedom Light, Abbott Laboratories, Lake Bluff, IL, United States)
ELISA assays for various biomarkers
Fast protein liquid chromatography (FPLC) with an AKTA apparatus (Pharmacia, Roosendaal, Netherlands) for lipoprotein profiling
Enzymatic assays (Roche diagnostics, Basel, CHF) for measuring total cholesterol and triacylglycerols in lipoprotein fractions

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