Simultaneous measurement of nineteen binding constants of peptides to vancomycin using affinity capillary electrophoresis-mass spectrometry.
On-line affinity capillary electrophoresis-electrospray ionization-mass spectrometry (ACE-MS) was used for the simultaneous measurement of multiple binding constants of an all-D-tetrapeptide library to the model receptor, vancomycin. Determination of Kd values for the 19 peptides of the form Fmoc-DXYA is demonstrated. The data are compared with the results obtained for individual compounds using ACE-UV, and good correlation between the two detection methods is shown. Simultaneous determination of multiple Kd values by ACE-MS is achieved in one set of experiments, whereas only one Kd value can be obtained by ACE-UV during the same time. ACE-MS measures multiple binding constants in solution in a fast and reliable manner using femtomole amounts of samples.
Dunayevskiy
Y M
YM
Barnett Institute, Northeastern University, Boston, Massachusetts 02115, USA.
Lyubarskaya
Y V
YV
Chu
Y H
YH
Vouros
P
P
Karger
B L
BL
D001667:Binding, Competitive; D019075:Electrophoresis, Capillary / Q000379:methods; D013058:Mass Spectrometry / Q000379:methods; D008956:Models, Chemical; D010455:Peptides / Q000737:chemistry*; D014640:Vancomycin / Q000737:chemistry*
D010455:Peptides; D014640:Vancomycin