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
