High-performance liquid chromatography of amino acids, peptides and proteins. LXXXVIII. Calculation of the average distance between protein solutes and the stationary phase during isocratic anion-exchange chromatography.
This investigation deals with protein retention behaviour in high-performance anion-exchange chromatography in terms of the average distance of approach between the protein solute and the positively charged anion-exchange stationary-phase surface. The theoretical treatment is based on a modified Debye-Hückel theory for spherical impenetrable ions, where the electrostatic potential energy has been related to the chromatographic capacity factor, k'. Results are presented for three globular proteins, eluted isocratically from a Mono-Q strong anion-exchange resin with sodium chloride as the displacer salt by a mobile phase with pH in the range 5.50-9.60. Analysis of experimental retention data indicates that topographically predefined, charged regions on the protein surface, called ionotopes, control the orientation and approach distance of the protein solute.
Hearn
M T
MT
Department of Biochemistry, Monash University, Clayton, Victoria, Australia.
Hodder
A N
AN
Aguilar
M I
MI
D000596:Amino Acids / Q000032:analysis*; D002852:Chromatography, Ion Exchange / Q000379:methods; D006863:Hydrogen-Ion Concentration; D008433:Mathematics; D010455:Peptides / Q000032:analysis*; D011506:Proteins / Q000032:analysis*; D013056:Spectrophotometry, Ultraviolet
D000596:Amino Acids; D010455:Peptides; D011506:Proteins