BCH 4123 Lecture Notes - Lecture 2: Nephelometer, Hyperlipidemia, Chemiluminescence
13 Jan 2017
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Enzymatic: glucose assay looks for increase in absorbance of nadh. Short wavelength so less interference (likely only lipemia) Dye binding assay: albumin measured by bcp or bcg. A complex is absorbed at 600nm: ca and mg measured in similar way. Nephelometry and turbidimetry: difference is that turbidimetry measures incidence light, nephelometry measures increase in scattered light. Can design system to interact specifically with different materials. Can tune the system to ignore effects of lipemia or other light scatterers. Uses a laser: hook /prozone effect when there is no wash between antibody/antigen binding. Ratio between ab and ag = gives maximum binding. Once ab is saturated, not as many macromolecules formed and less scattering of light; rapid decrease in signal (hook/prozone effect) Can misread the concentration of the analyte. Fpia: fluorescent molecule depolarizes light, bind a receptor to increase mass of molecule to reduce depolarization, measure the quantity by the change in light polarization.
