22MOBT - Selected bioanalytical techniques
| Course specification | ||||
|---|---|---|---|---|
| Course title | Selected bioanalytical techniques | |||
| Acronym | 22MOBT | |||
| Study programme | Biochemical Engineering and Biotechnology | |||
| Module | ||||
| Lecturer (for classes) | ||||
| Lecturer/Associate (for practice) | ||||
| Lecturer/Associate (for OTC) | ||||
| ESPB | 4.0 | Status | ||
| Condition | Облик условљености | |||
| The goal | The 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 biotechnological processes or being products of the same, such as proteins, carbohydrates, lipids, nucleic acids and vitamins. | |||
| The outcome | Students 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 for determination of concentration in commercial products or in following the course of particular bioprocesses, such as enzymatic reactions, fermentations and enzyme immobilizations. | |||
| Contents | ||||
| Contents of lectures | Chromatography 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. Quantitation assays of biomolecules. Protein quantitation assays. Carbohydrate quantitation assays. Nucleic acid quantitation assays. Lipid quantitation assays. Vitamin quantitation 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. Statistical analysis. Precision and accuracy. Limits of quantitation and detection. | |||
| Contents of exercises | Spectroscopic assays for total sugars and reducing sugars. Determination of mono-, di- and trisaccharide concentration using high pressure liquid chromatography (HPLC) system equppied with sugar columns (amino-functionalyzed columns or columns with immobilized metal counterions for formation of chelate complexes with sugars) and refractive index detector. Determination of hydrosoluble and liposoluble vitamins using reversed phase HPLC in pharmaceuticals and nutraceuticals. Colorimetric protein assays. Monitoring the course of enzyme-catalyzed protein hydrolysis with ninhydrin assay and reversed-phase HPLC. Validation of bioanalytical method for determination of enzymatic activity. Statistical analysis of accuracy and precision (LOD and LOQ, recovery test, inter-laboratory precission). | |||
| Literature | ||||
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| Number of hours per week during the semester/trimester/year | ||||
| Lectures | Exercises | OTC | Study and Research | Other classes |
| 1 | 2 | |||
| Methods of teaching | Theoretical lectures. Laboratory practice. | |||
| Knowledge score (maximum points 100) | ||||
| Pre obligations | Points | Final exam | Points | |
| Activites during lectures | Test paper | |||
| Practical lessons | 40 | Oral examination | 60 | |
| Projects | ||||
| Colloquia | ||||
| Seminars | ||||