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Protein Purification 3: Anion Exchange Chromatography

This resource addresses these Core Concepts of Vision and Change: 1. EVOLUTION 2. STRUCTURE AND FUNCTION 4. PATHWAYS AND TRANSFORMATIONS OF ENERGY AND MATTER This resource can help students develop these Core Competencies of Vision and Change: 1. ABILITY TO APPLY THE PROCESS OF SCIENCE 2. ABILITY TO USE QUANTITATIVE REASONING 5. ABILITY TO COMMUNICATE AND COLLABORATE WITH OTHER DISCIPLINES These materials are intended to be part of a Vision and Change Collection about enzyme purification. This lab activity completes the partial purification of adenosine deaminase (ADA). Dialyzed fractions from size exclusion chromatography are loaded onto anion exchange columns, and a salt gradient is used to elute ADA. Enzyme activity is located using a QUALITATIVE spectrophotometric assay, and pooled active fractions are measured at 280 nm. All the measurements for an enzyme purification table are completed, including QUANTITATIVE activity assays for each purification fraction. Student teams complete the objectives and analyze the data. The pre-lab lecture component uses SCALE-UP to engage the students and prepare them for lab. Students watch “lecture chunks” (created by breaking up my traditional 60 minute Ppt lectures into 3-5 minute video clips) before the pre-lab meeting: these short presentations about biochemical theory and lab set-up allow the instructor to “flip the classroom” (i.e., learn before lecture) so class time can incorporate more student-centered activities. Additional videos about chromatography station set-up and loading samples are included in "Protein Purification 3: Anion Exchange Chromatography Supplemental." I use additional Ppt slides with clicker questions during the class discussion before lab that complement the student-centered activities. These are tested materials currently used in a seven day advanced undergraduate laboratory course (BIOC 311: Advanced Experimental Biosciences) that is targeted at juniors and seniors.

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Resource Type: Digital Presentation (Powerpoint), Short Movies, Assessment: other, Laboratory or Hands-On Activity, Laboratory Manual, Lecture / Lecture Outline / Lecture Notes, Teaching Strategies & Guidelines, Tutorial
Audience Level: Undergraduate upper division (Grades 15-16), Graduate

Author and Copyright

Authors and Editors: PhD Beth Beason-Abmayr of Rice University Biochemistry and Cell Biology
Publisher: Rice University Biochemistry and Cell Biology
Copyright and other restrictions: No
Cost: No


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