The Biochemical Techniques laboratory course at Santa Clara University features a multi-week project in protein purification and characterization. The protocol allows student teams to isolate three different proteins from a single sample of tissue from a nontraditional source, mammalian blood, via a series of successive column chromatographic procedures.  By use of a microplate reader student teams are able to assess the relative protein concentration and enzyme activity in a large number of chromatography column fractions in a relatively short period of time.  Students are able to quickly perform a real-time critical evaluation of their data to optimize the preparation of starting material for subsequent steps in the purification scheme.  The protocol consistently results in a high degree of fold purification and a relatively high recovery of activity for each protein.   Students gain experience in processing large quantities of experimental data in a timely fashion, in working in a team environment, and in carrying out a variety of different biochemical techniques such as column chromatographic separations, microscale assays, and native and SDS polyacrylamide gel electrophoresis.  The protocol may also be modified in a variety of ways to serve as a springboard for research-style individual or small-group projects during the second half of the term.

Development of a multi-week protein purification and characterization project lab for upper division biochemisty and biotechnology laboratory courses

248th National Meeting (2014)

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