M. Weitzhandler et al., Protein variant separations by cation-exchange chromatography on tentacle-type polymeric stationary phases, J CHROMAT A, 828(1-2), 1998, pp. 365-372
We developed a set of prototype cation-exchange column packings that are ba
sed on a hydrophilic coated, pellicular polymeric support with a grafted te
ntacular surface chemistry that is highly suited to resolving closely relat
ed protein variants. These column packings (1) afford minimal band spreadin
g in conjunction with extremely high selectivity, (2) exhibit a very hydrop
hilic character and (3) have moderate loading capacity. Cytochrome c varian
ts (bovine, horse, rabbit) were baseline-separated, as was native ribonucle
ase A and its two deamidation products, the Asp(67) and isoAsp(67) forms. H
umanized monoclonal antibody variants differing in the presence of lysine a
t the C terminus of the heavy chains were baseline-resolved. Finally, the s
eparation of hemoglobin variants found in a sample containing elevated leve
ls of glycated hemoglobin was also demonstrated. (C) 1998 Published by Else
vier Science B.V. All rights reserved.