Lectures 10 14: Membrane Filtration and Dialysis

1 Lectures 1 – 9: Spectroscopy

Introduction, absorption spectroscopy and electromagnetic spectrum. Chromophore and energy level. Beer’s law and its deviations, spectrophotometer and colorimeter. Parameters measured in absorption properties of a chromophore. Applications of absorption spectroscopy. Reporter groups.

Lectures 10 – 14: Membrane Filtration and Dialysis

Nitrocellulose filters and membrane filtration, fiberglass filters. Applications of nitrocellulose and fiberglass filters. Polycarbonate filters and their uses.

Dialysis and membrane filtration. Factors affecting the rate of dialysis. Reverse dialysis, glass fiber dialysis. Molecular filtration – diaflo and pellicon.

Lectures 15 – 26: Chromatography

Partition chromatography: theory, uses and examples.

Adsorption chromatography: Types of adsorption chromatography.

Operation of columns, elution, supports

Paper chromatography: detection and identification of spots.

Thin layer chromatography

Gas liquid chromatography: its advantages and uses.

Gel chromatography: theory, materials used, advantages and applications.

Thin layer gel chromatography and its uses.

High performance liquid chromatography and its applications

Ion exchange chromatography: principles, properties of ion exchangers, choice of ion exchangers, porosity and mesh size, capacity, choice of pH, buffer and ionic conditions. Applications of ion exchange chromatography.

Affinity chromatography and its applications.

Hydroxyapatite chromatography.

Lectures 27 – 32: Hydrodynamic methods

Sedimentation, simple theory of velocity-sedimentation. Instrumentation for ultracentrifugation. Analytical and preparative ultracentrifuge. Boundary- sedimentation-velocity experiment. Factors affecting sedimentation velocity.

2 Zonal centrifugation through reformed density gradient. Determination of molecular weight by sedimentation.

Lectures 33 – 37: Electrophoresis and Isoelectric focusing

Theory of electrophoresis, types of electrophoresis: moving boundary and zone electrophoresis, paper electrophoresis, cellulose acetate strip and polyacrylamide gel electrophoresis,. SDS-PAGE- vertical and horizontal electrophoresis and their applications. Agarose gel electrophoresis and its applications. Isoelectric focusing its theory and applications. Fingerprinting and large-scale preparative electrophoresis.

Lectures 38 – 41: Radioisotope Techniques

Nature of radioactivity (atomic stability and radiation, types of radioactive decay). Detection and measurement of radioactivity. Radioactive counting. Applications of radioisotopes in biochemistry. Radiation risk and safety.

Lectures 42 -44: Immunological Methods

Immunological system. Preparation of antibodies and immune reaction. Immunodiffusion methods. Radioimmunoassay. Immunoradiometric assay and immunoelectrophoresis.

Books:

Physical Biochemistry: Principles and Applications by David Sheehen

Physical Biochemistry: Applications to Biochemistry and Molecular Biology by David M. Friefelder.

Practical Biochemistry – Principles and techniques by Keith Wilson and John Walker