Hydrophobic interaction (HI) chromatography

HI-chromatography was discovered serendipitously  when, in control experiments,  ligands  were  omitted  from  the  matrix/spacer  arm combination.  It  was  found  that  the  resulting  resins  were  nevertheless effective at separating proteins, due to  hydrophobic  interactions  between the  sample  proteins  and  the  aliphatic  spacer  arms.  Following  this discovery,  HI-resins  were  purposefully  designed  to  optimize  the hydrophobic  interaction.

Hydrophobic “bonds” are increased in strength by an increase in buffer ionic strength.  HI-chromatography  therefore  conveniently  fits  into  an isolation  scheme,  immediately  after  a  salting  out  step,  as  the  high  salt levels will promote binding to the HI-resin.  Proteins  can subsequently be eluted by decreasing the buffer ionic strength, either in a stepwise manner or in a gradient.

References

-Dennison, C. (2002). A guide to protein isolation . School of Molecular mid Cellular Biosciences, University of Natal . Kluwer Academic Publishers new york, Boston, Dordrecht, London, Moscow .