Using an FPLC to promote active learning of the principles of protein structure and purification

Abstract

The concepts of protein purification are often taught in undergraduate biology and biochemistry lectures and reinforced during laboratory exercises; however, very few reported activities allow students to directly gain experience using modern protein purification instruments, such as Fast Protein Liquid Chromatography (FPLC). This laboratory exercise uses size exclusion chromatography (SEC) and ion exchange (IEX) chromatography to separate a mixture of four different proteins. Students use an SEC chromatogram and corresponding SDS-PAGE gel to understand how protein conformations change under different conditions (i.e. native and nonnative). Students explore strategies to separate co-eluting proteins by IEX chromatography. Using either cation or anion exchange, one protein is bound to the column while the other is collected in the flow-through. In this exercise, undergraduate students gain hands-on experience with experimental design, buffer and sample preparation, and implementation of instrumentation that is commonly used by experienced researchers while learning and applying the fundamental concepts of protein structure, protein purification, and SDS-PAGE. V C 2016 by The International Union of Biochemistry and Molecular Biology, 45(1):60-68, 2017.

Exploring DNA in biochemistry lab courses: DNA barcoding and phylogenetic analysis

“…Students need to master methods of nucleic acid isolation, purification, and learn PCR in addition to basics of DNA sequencing and bioinformatics. 3,4 This knowledge can be enforced through the hands-on laboratory exercises to allow students to gain skills necessary to succeed in either academic research or industrial environment. One of the methods that combine all the listed techniques is DNA barcoding.…”

“…Undergraduate biology and biochemistry courses curricula cover the fundamentals of DNA structure and function, 1,2 as well as experimental techniques based on the unique informational features DNA holds. Students need to master methods of nucleic acid isolation, purification, and learn PCR in addition to basics of DNA sequencing and bioinformatics 3,4 . This knowledge can be enforced through the hands‐on laboratory exercises to allow students to gain skills necessary to succeed in either academic research or industrial environment.…”