BIOCHEMISTRY AND CLINICAL BIOCHEMISTRY
Learning outcomes of the course unit
The course as its formative aspective to provide a basic knowledge of the analytical techniques used in the biochemical and clinical biochemistry laboratories.
Course contents summary
Centrifugation - Sedimentation theory. Calculations of centrifugal force and centrifugation times. Homogenisation: omogenizers and omogenization media. Types of centrifugal separations: differential centrifugation, grade centrifugation, rate zonal centrifugation, isopicnic centrifugation. Types of centrifuge. Centrifuge rotors.
Chromatography - Retention in chromatography. Distribution coefficient. Classification of chromatographic techniques. Peak shape. Sorption isotherms. Capacity factor. Column efficiency. Selectivity and resolution. Band broadening processes. Separation modes: absorption chromatography, partition chromatography, bonded phases: reverse-phase chromatography, ion-exchange chromatography, ion-pair chromatography, size exclusion chromatography, affinity chromatography. High performance liquid chromatography (HPLC): pumps, valve injection, columns, detectors.
UV-VIS absorption spectroscopy - Properties of electromagnetic radiation. Interaction electromagnetic radiation-matter. Chromophores and transitions. Absorption intensity. Selection rules. Spectrophotometers. Quantitative and qualitative analysis.
Radioactivity and its measurement - Types of radioactive emission. Mode of radioactive decay. Interaction of radiations with matter. Detection and measurement of radioactivity: ionization chambers, solid scintillation counting, liquid scintillation counting.
Radioimmunoassay (RIA) and Enzymeimmunoassay (EIA) - Antigen and antibody. Labeled antigen. Antigen-antibody reactions. Principle of radioimmuno assay. EIA: antigen labelled with enzyme. General features of quantitative radio- or enzyme immunoassay.
frontal lesson with slides