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14MOBT - Selected bioanalytical techniques

Course specification
Course titleSelected bioanalytical techniques
Acronym14MOBT
Study programme
Module
Lecturer (for classes)
Lecturer/Associate (for practice)
    Lecturer/Associate (for OTC)
      ESPB4.0Status
      ConditionОблик условљености
      The goalThe main goal is to enable students to use the most important methods and techniques for identification, quantification and characterization of biomolecules. The most relevant methods for quantitative determination of biomolecules, used as substrates of biochemical processes or being products of the same, such as proteins, carbohydrates, lipids, nucleic acids and vitamins.
      The outcomeStudents will learn chromatographic and spectroscopic methods applied in quantitative analysis of biomolecules, as well as electrophoretic methods of biomolecules characterization. During the laboratory work these methods will be applied in following the course of particular bioprocesses, such as enzymatic reactions, fermentations and enzyme immobilizations.
      Contents
      Contents of lecturesChromatography of biomolecules. Plate theory of chromatography. van Deemter equation. Detectors and elution regimes. Ion exchange chromatography. Affinity chromatography. Gel permeation chromatography. Reversed-phase chromatography. Hydrophobic interaction chromatography. ; Quantification assays for biomolecules. Protein quantification assays. Carbohydrate quantification assays. Nucleic acid quantification assays. Lipid quantification assays. Vitamin quantification assays. ; Electrophoretic characterization of biomolecules. Electrophoretic support media. Detection of proteins and nucleic acids after electrophoretic separation. Zone electrophoresis. Isoelectric focusing. ; Validation of new bioanalytical methods. Precision and accuracy. Limits of quantitation and detection. ;
      Contents of exercisesSpectroscopic assays for total sugars and reducing sugars. Glucose determination with spectroscopic assay using GOD-PAP reagent. Application of specific method for glucose determination in following the course of enzyme-catalyzes lactose hydrolysis and glucose isomerisation. Chromatografic determination of monosaccharides and oligosaccharides. ; Protein and nucleic acid assays. Following the course of protein immobilization and microbial enzyme production and electrophoretic characterization of final product. ; Chromatographic determination of vitamins and physiologically active compounds. Chromatographic determination of fatty acid composition of fats and oils. ;
      Literature
      1. Mikkelsen S. R., Corton E, Bioanalytical chemistry, Wiley-Interscience, 2004.
      2. Rosenberg I.M., Protein analysis and purification. Benchtop techniques, Birkhäuser, 1996.
      3. Hames B.D., Gel electrophoresis of proteins: A practical approach, Oxford University Press, 1998.
      4. Kromidas S., Practical problem solving in HPLC, Wiley-VCH, 2000.
      Number of hours per week during the semester/trimester/year
      LecturesExercisesOTCStudy and ResearchOther classes
      12
      Methods of teachingTheoretical lectures. Laboratory practice. Problem solving and calculus.
      Knowledge score (maximum points 100)
      Pre obligationsPointsFinal examPoints
      Activites during lecturesTest paper40
      Practical lessonsOral examination
      Projects
      Colloquia40
      Seminars20