BIOT 233 - Protein Analysis and Purification

PREREQUISITES: BIOT 212 - Analytic Methods in Biotechnology II
PROGRAM: Biotechnology
CREDIT HOURS MIN: 4
LECTURE HOURS MIN: 2
LAB HOURS MIN: 4
DATE OF LAST REVISION: Summer, 2011

Students will review the biochemical properties of amino acids and proteins, then study techniques of cell disintegration and extraction, protein separation, and analysis. Students will be taught to determine which method is most applicable in various situations and why that method should be utilized. When possible, students will be given an opportunity to perform these techniques in the laboratory.

MAJOR COURSE LEARNING OBJECTIVES: Upon successful completion of this course the student will be expected to:

Describe the function and structure of proteins.
Be able to understand the principles of chromatography separation techniques involved in protein purification and be able to apply these to a given protocol.
Be able to run spectrophotometric assays to determine protein concentration of a sample.
Be able to separate proteins by gel electrophoresis.
Describe the principles of enzyme assays and be able to determine the activity of an enzyme based on spectrophotometric assays.
Know the principles involved in protein detection/measurement through ELISA and Western blot.
Understand principles of protein engineering and expression.

COURSE CONTENT: Topical areas of study include -

Overview of protein biotechnology and proteomics
Structure and function of proteins
Protein synthesis and post-translational modifications
Protein sequencing and amino acid analysis
Principles of spectrophotometry and centrifugation
Techniques of salting out, organic solvent precipitation, selective denaturation
Techniques of ultrafiltration, dialysis, and lyophilization
Protein assay and quantification
Protein chromatography (SDS-PAGE, HPLC, FPLC, SEC, affinity, ion exchange, reversed phase)
Protein bioinformatics and structure determinations
Industrial protein production and scale-up
Enzymes and enzyme activity
ELISAs
Western blots
Proteomics and protein microarray
Instrumentation including pH meter, centrifuge, spectrophotometer, fraction collector, electrophoresis equipment, tissue lysis and homogenizer

Course Addendum - Syllabus (Click to expand)

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BIOT 233 - Protein Analysis and Purification