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Purifying Challenging Proteins—Principles and Methods

The Purifying Challenging Proteins Handbook is intended for students and experienced researchers with an interest in the isolation of integral membrane proteins, multiprotein complexes, or in refolding proteins from inclusion bodies.

Our knowledge and understanding of the structural and functional biology of soluble proteins has increased dramatically over the last decade. Much of the technology for the production, purification, and analysis of soluble proteins is now at a stage where generic purification protocols allow relatively high success rates.

The situation is different for integral membrane proteins, multiprotein complexes, and inclusion bodies. The need to handle and study these more difficult groups of proteins is clear, given that:

about 30% of a typical cell’s proteins are membrane proteins, and more than 50% of the current drugs on the market exert their actions via membrane proteins
while carrying out their enzymatic, structural, transporting, or regulatory functions, proteins most often interact with each other, forming multiprotein complexes
a large proportion of normally soluble proteins that are overexpressed in E. coli end up as incorrectly folded and insoluble protein in inclusion bodies

The aim of the handbook is to present tools, strategies, and solutions available to meet the purification challenges associated with these three classes of proteins.

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