Oncology 675 - Protein Purification

This is a 2-credit lecture course that meets from 1:20-2:15 PM on Tuesdays and Thursdays during the Fall semester (odd-numbered years), in room 125 McArdle. It will be given next in Fall 2005. It is primarily a lecture course consisting of 27 lectures, 2 half-semester exams, some take-home problems, and a paper on a topic relating to the course. The students are asked to read at least twelve articles and prepare an informative summary with references. Students are expected to read "Protein Purification; Principles and Practice" Scopes, 3rd edition.

Goals of the course are: 1) to introduce the most important and useful concepts of protein purification and handling, 2) to help students to develop an intuition about how to work with proteins -- so that they can "think like a protein", and 3) to guide students to ongoing sources of information and resources.

Lecture topics include : Introduction-Protein purification overview; Properties of proteins/types of separation methods; Assays -- following an enzyme through a purification; Protein characterization; Protein inactivation and stabilization/solution components; Purification strategy/starting materials/preparing cell extracts; Precipitation methods; Computer simulation of protein purification; Phase partitioning; Dialysis, desalting, concentration, and ultrafiltration; Preparative electrophoresis, chromatofocusing, isoelectric focusing, capillary electrophoresis; Purification of membrane proteins/glycoproteins; Column chromatography -- theory and concepts; Sizing -- gel filtration chromatography; Ion exchange, Affinity, Immunoaffinity, and DNA affinity chromatography; HPLC: Columns and hardware, theory, methods development, applications; Micropurification by eluting from SDS gels; Overproduction of cloned gene products; Purification of insoluble overproduced proteins; Engineering proteins for ease of purification and characterization; Recent advances in studying protein-protein interactions; Immobilizing and using enzymes in bioreactors; Scale-up considerations/related resources, courses, and facilities.

This course is usually taken for credit by about 25-30 graduate students, and occasionally by senior undergraduates. Auditors or those who wish to sit in on part or all of the lectures are welcome (usually as many as 20-30 specialists, graduate students, postdocs, and faculty sit in).
For more info contact: Dick Burgess - burgess@oncology.wisc.edu