Use of affinity capillary electrophoresis for the study of protein and drug interactions.
Protein-drug interactions were studied using affinity capillary electrophoresis (ACE). The initial study was performed using a model system, fibronectin-heparin interaction. Two distinct binding constants, 21 and 641 nM, were derived from the Scatchard plots. The results are consistent with reported data obtained using other analytical techniques. The ACE binding assay was applied for studying molecular interactions between kedarcidin chromophore and apoprotein. Conditions with an organic solvent as buffer component were examined to establish a suitable binding assay. It appears that the electrophoretic behavior of the protein shows little distortion in the presence of either dimethyl sulfoxide (up to 10%) or acetonitrile (ACN) (up to 30%). The binding assay was initially conducted in aqueous buffer phase. The saturation concentration of chomophore was found to be around 15 microM. A linear Scatchard plot was derived from binding data with a correlation coefficient of 0.94. The binding constant was determined as Kd = 5.6 microM. The effects of organic solvent content ranging from 0 to 30% ACN on the constant were examined. The binding constants were determined as Kd = 11, 12.5 and 16.2 microM for 5, 10 and 30% ACN, respectively. It appeared that the binding affinity between kedarcidin chromophore and apoprotein is reduced as the organic solvent content in the aqueous phase is increased.
Liu
J
J
Bristol-Myers Squibb Pharmaceutical Research Institute, Princeton, NJ 08543, USA.
Abid
S
S
Hail
M E
ME
Lee
M S
MS
Hangeland
J
J
Zein
N
N
D000480:Alkynes / Q000378:metabolism*; D000818:Animals; D001059:Apoproteins / Q000378:metabolism*; D003516:Cycloparaffins / Q000378:metabolism*; D004347:Drug Interactions; D019075:Electrophoresis, Capillary; D053281:Enediynes; D006493:Heparin / Q000378:metabolism*; D009281:Naphthalenes / Q000378:metabolism*
D000480:Alkynes; D001059:Apoproteins; D003516:Cycloparaffins; D053281:Enediynes; D009281:Naphthalenes; C080542:kedarcidin chromophore; D006493:Heparin