This paper describes an undergraduate
biochemistry laboratory module
consisting of a set of experiments designed around a purification
scheme for bovine serum albumin (BSA). Students purify BSA from cow
plasma by a combination of salt and acetone precipitation, equilibrium
dialysis, ion exchange, and size exclusion chromatography. Students
use the bromocresol green–BSA complex assay to quantify albumin
at each step in the scheme and generate a purification table. In addition,
trace amounts of IgG’s and nucleases, the major contaminants
in BSA purification, are probed by Western blotting and DNase assay,
respectively. This module exposes students to the principles and techniques
of separation of proteins based on solubility, size, and surface charge.
In addition, students learn the concept of limit of detection of analytical
techniques such as SDS-PAGE and Western blotting. The module utilizes
two 3-h laboratory periods, with a 1-h prelab lecture per week, for
about 6 weeks. Depending on class size, groups consist of two to four
students. The module is suitable for students who have completed a
semester of biochemistry. Instructors can extend the module to advanced
level laboratory by including enzyme kinetics and protein structure–function
studies based on albumin’s pseudoesterase activity and intrinsic
tryptophan fluorescence, respectively.
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