Hydrophobic Interaction and Reverse Phase

0 downloads 0 Views 373KB Size Report
Hydrophobic Interaction. Chromatography(HIC). Hydrophobic Interaction and Reverse Phase. Chromatographies. THE 2ND SUMMER SCHOOL IN.
P7

Hydrophobic Interaction and Reverse Phase Chromatographies

THE 2ND SUMMER SCHOOL IN INTERDISCIPLINARY SCIENCES 21-24 september 2009

Presenter: Z. Behzadian 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

Hydrophobic Interaction Chromatography(HIC)

Stationary phase of HIC

HIC is temperature sensitive

Hydrophobic residues of proteins usually are buried internally. The number and distribution of hydrophobic residues are varied on the surface of proteins which can be used in HIC. When a hydrophobic region of a biopolymer binds to the surface of a mildly hydrophobic stationary phase, hydrophilic water molecules are effectively released from the surrounding hydrophobic areas causing a thermodynamically favorable change in entropy.

Reverse- phase chromatography (RPC)

Temperature and ionic strength play strong roles in HIC.

Reverse- phase chromatography includes any chromatographic method that uses a non-plar stationary phase.

Ammonium sulfate, by virtue of its good salting-out properties and high solubility in water is used as an eluting buffer

Principles of RPC

Hydroponic interactions is ionic strength sensitive Elution of bind protein in HIC

Hydrophobic intraction chromatography (ligand/protein “salting out)

Stationary phases for reversed-phase chromatography