LIPIDS: Lipoproteins II
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Major protein found in HDL ; activates LCAT; ANTIATHEROGENIC
Apo A1
Major Protein found in LDL ; Risk of CHD
Apo B100
Independent risk factor ; Impaired Plasminogen Activation
Lipoprotein a
LABORATORY METHODS FOR LIPOPOROTEINS
Ultracentrifugation Electrophoresis
Laboratory Method for Lipoproteins: Based on Density and Shape Sample is adjusted with Sodium Bromide and is centrifuged at high speed for 24 HOURS
Ultracentrifugation
Laboratory Method for Lipoproteins: Reference method for Lipoproteins
Ultracentrifugation
Laboratory Method for Lipoproteins: Order from MOST TO LEAST DENSE (ULTRACENTRIFUGATION)
• HDL (bottom) • LDL • VLDL • Chylomicrons (Top;creamy layer)
Laboratory Method for Lipoproteins: Separation based on size and charge
Electrophoresis
Laboratory Method for Lipoproteins: Stains for Electrophoresis for Lipoprotein
Oil Red O Sudan Black B Sudan III Sudan IV Fat Red B
Laboratory Method for Lipoproteins: Arrangement from fastest/most anodic (Electrophoresis)
HDL > LDL > VLDL > CM
Laboratory Method for Lipoproteins: Test primarily used for Chylomicrons; Involves standing aliquot of plasma in Refrigerator for 4 deg celsius overnight
Standing Plasma Test
Laboratory Method for Lipoproteins: Standing Plasma Test: The aliquot remains turbid even after standing overnight
Excess BVLDL
Laboratory Method for Lipoproteins: Standing Plasma Test: Presence of Chylomicrons at a Fasting Plasma
Abnormal
Laboratory Method for Lipoproteins: Polyanion Precipitation (3-step Precipitation Method) Homogenous Assay These test are used for the measurement of ?
HDL Measurement
Laboratory Method for Lipoproteins: In Polyanion Precipitation (3-step Precipitation Method): Agents used to precipitate Lipoprotein with Apo B
Dextran Sulfate Heparin Sulfate Phosphotungstate Divalent cation: Magnesium Calcium Manganese
Laboratory Method for Lipoproteins: In Polyanion Precipitation (3-step Precipitation Method): Reagents used
Heparin sulfate - Manganese Chloride Dextran Sulfate - Magnesium Chloride
Laboratory Method for Lipoproteins: In Polyanion Precipitation (3-step Precipitation Method): HDL supernatant is measured using (2)
Cholesterol Oxidase Cholesterol esterase
Laboratory Method for Lipoproteins: In Polyanion Precipitation (3-step Precipitation Method): Can be automated T/F
FALSE POLYANION CANNOT BE AUTOMATED
Laboratory Method for Lipoproteins: Used for HDL measurement; uses antibody to bind with Apo B100 (VLDL, LDL) HDL then is measured ENZYMATICALLY Principle is used in automated measurement
Homogenous Assay
Laboratory Method for Lipoproteins: • Friedawald Equation • Delong Equation • Beta Quantification • Homogenous Direct LDL Method • Gel Chromatography / Affinity Chromatography • Immunochemical • Immunoassay or immunonephelometry
LDL Measurement
Laboratory Method for Lipoproteins: LDL Measurement suitable if TRG is <400
Friedwald Equation
Laboratory Method for Lipoproteins: LDL = TC - (HDL + VLDL) VLDL = TRG / 5 (g/dL) VLDL = TRG / 2.175 (mmol/L)
Friedwald Equation
Laboratory Method for Lipoproteins: LDL measurement used when TRG is >400 g/dl
Delong Equation
Laboratory Method for Lipoproteins: LDL = TC - (HDL + VLDL) VLDL = TRG / 6.5 (g/dL) VLDL = TRG / 2.825 (mmol/L)
Delong Equation