BCH 332 Biophysical Biochemistry

  • Days: Sunday, Tuesday, and Thursday
  • Building: 5
  • Room:
  • Time: 9.00 - 9.50 AM
  • Course Symbol: BCH 332
  • Course titleBiophysical Biochemistry
  • Credit hours: 3 (3+0)
  • Prerequisite: BCH 312

 

  • A course designed to study the methods for purification and characterization of biomolecules.  The topics of this course include biochemical applications of Spectroscopy (absorption, fluorescence, and Mass spectroscopy), basic & common methods (tissue homogenization, dialysis, filtration, and salting out), Hydrodynamic methods (various forms and applications of centrifugation), Electrophoresis (paper, PAGE, and agarose), various forms and applications of Chromatography (gel filtration, ion-exchange, adsorption, affinity, and HLPC), and Radioisotope applications in Biochemistry.
  • Lectures:

    1. Lectures 1-5: Basis of electromagnetic spectrum and energy levels.  Absorption spectroscopy and chromophores. Beer-Lambert law and application of absorption spectroscopy (protein determination, nucleic acid, enzyme kinetics).
    2. Lectures 6 -7: Fluorescence spectroscopy.
    3. Lecture 8: Mass spectroscopy.
    4. Lecture 9: Various methods of tissue homogenization.
    5. Lecture 10: Stabilization of samples (inhibitors, chelators, glycerol etc)
    6. Lecture 11: Salting in and salting out, protein precipitation.
    7. Lecture 12: Organic solvent extraction.
    8. Lectures 13 – 14: Dialysis, filtration, pressure dialysis, common filters.
    9. Lectures 15 – 16: Basic concept of sedimentation (Svedberg equation).
    10. Lecture 17: Analytical and preparative centrifuges (types and rotors).
    11. Lecture 18 – 19: Applications of centrifugations: cell fractionation, sucrose density gradient, CsCl gradient and analytical applications.
    12. Lecture 20: Theory and types of electrophoresis.
    13. Lecture 21: Strip electrophoresis and its applications.
    14. Lectures 22 - 24: Polyacrylamide gel electrophoresis (native, SDS-PAGE and isoelectric focusing).
    15. Lecture 25 Agarose gel electrophoresis (analytical and preparative applications).
    16. Lecture 26: Gel visualization methods (coomasive blue, silver staining, ethidium bromide etc.).
    17. Lecture 27: Basic concepts and theory of chromatography.
    18. Lecture 28: Classification of chromatography (gas, liquid, thin layer etc).
    19. Lecture 29: Gel filtration.
    20. Lecture 30: Ion exchange chromatography.
    21. Lecture 31: Adsorption chromatography.
    22. Lecture 32: Hydrophobic chromatography.
    23. Lecture 33: Affinity chromatography.
    24. Lecture 34: High pressure liquid chromatography.
    25. Lectures 35- 36: Immunological methods – basic concepts, preparative applications and analytical applications. ELISA, RIA, diffusion assays.
    26. Lectures 37 - 42: Radioisotopes – basic concepts, units of radioactivity, radioactive decay and half life, commonly used isotopes in biochemistry, radioisotope applications in biochemistry, detection and measurement of radioactivity (Geiger-Muller counter, scintillation counter and film exposure).

 

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