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Ion Exchange Chromatography

Poster · June 2016

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Reza Yousefi
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P4

Ion-Exchange Chromatography
THE 2ND SUMMER SCHOOL
IN INTERDISCIPLINARY
SCIENCES
21-24 September 2009

Presenter:M.Taheri
Supervised by Dr.R.Yousefi (Assistant Professor of Biochemistry)
Co- Supervisor: Dr.H.R.Karbalaei (Assistant Professor of Biochemistry)
Department of Biology,College of Sience,Shiraz University,Shiraz 71454,Iran

Ion Exchange Some biochemically useful ion exchangers Stepwise / Linear Elution
Chromatography (IEC)
Ion-exchange chromatography (or ion
chromatography) is a process that allows the
separation of ions and polar molecules based
on their charge. It can be used for almost any
kind of charged molecule including large
proteins, small nucleotides and amino acids.

IEC Techniques
Strong ion exchangers (like SP and
QAE) are ionized over a wide pH range
Weak ion exhangers (like DEAE or CM)
are useful over a limited pH range
Choice of resin/matrix depends on:
Principle is to separate on basis of – Scale of separation
charge “adsorption” – Molecular size of components
Positively charged proteins are – Isoelectric point of desired protein
reversibly adsorbed to immobilized – pH stability of the protein of interest
negatively charged beads/polymers
Selecting a resin and starting
Negatively charged proteins are
reversibly adsorbed to immobilized
positively charged beads/polymers
Schematic representation of cation
Important considerations
exchange chromatography

If a protein is most stable below its pI,

a cation exchanger should be used
If a protein is most stable above its pI,
an anion exchanger should be used
If stability of the protein is known to
be good over a wider pH range then
either type of ion exchanger can be Chromatofocusing
IEC Advantages used

Proteins bind to charged matrix in low salt

Has highest resolving power
Has highest loading capacity
Widespread applicability (almost
universal)
Most frequent chromatographic
technique for protein purification
Used in ~75% of all purifications

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