Simultaneous estimation of the association constants of glycoprotein glycoforms to a common protein by capillary electrophoresis

Separation, identification, and interaction of heparin oligosaccharides with granulocyte-colony stimulating factor using capillary electrophoresis and mass spectrometry

“…Among separation and interaction techniques, CE has been proved to be very attractive due to the advantages of high separation efficiency, great flexibility, simple operation, short analysis time, and low consumption of samples and buffers [9]. In addition, the interaction between molecules can be directly observed under physiological conditions, without prior immobilization steps, as is required in affinity chromatography and surface plasmon resonance spectrometry [28][29][30][31][32][33][34][35][36] that pose a problem of steric hindrance [37][38][39]. It would also be possible, by using CE, to study interactions of individual components in a mixture and to determine binding parameters simultaneously [40].…”

Search for serum protein‐binding disaccharides and disaccharide‐binding serum proteins by affinity capillary electrophoresis

“…Each of the resultant buffers were filled in a capillary, and the diluted serum was introduced to the capillary by suction for 1.5 s using the vacuum injection system of the CE apparatus. The migration time was measured accurately at the peak top position using an integrator and corrected as described in [13]. Fluctutaion of migration time is usually caused by the variation of EOF velocity, whereas m ep is almost unchanged throughout analytical runs, provided the analysis is performed in the same capillary using the same running buffer at a controlled temperature.…”

“…The reverse system affords the advantage that interactions between multiple ligands and a common protein can simultaneously be observed, because ACE allows concurrent separation of the ligands while each ligand is interacting with the protein. We have demonstrated this advantage also in the simultaneous estimation of the association constants of glycoprotein glycoforms toward a specified lectin [13]. Simultaneous observation of multiple interactions is also possible in the normal system, if sample contains multiple species of proteins that can be separated from each other during the binding study by ACE.…”

Capillary electrophoresis of proteins 2001–2003

“…CE in different formats including zone electrophoresis and frontal analysis was used to a number of binding studies and investigated binding of ribonuclease and ovalbumin to agglutinin from Lens culinaris [211], drugs to human serum albumin and a 1 -acid glycoprotein [131], teicoplanin and D-Ala-D-Ala terminated peptides [212], enediynes [213], fosfatidylserine [214], and heparin [215] to BSA, anionic drugs [216] and oxybutinin to plasma proteins [217], drugs to plasma lipoproteins [218] and to subdomain III of HSA [219], porfyrin to HSA [220,221], heptamethine cyanine dye to HSA [41], phosphates to lysozyme, lactoferrin, and a-lactoglobulin [222] DNA to proteins [81], basic drugs to human a 1 -acid glycoprotein [223], further interactions of pUC19DNA and ovalbumin [224], 3'-azido-3'-deoxythymidine [225], oxovanadium ions [226], and herbicides atrazine and 2,4-dichlorophenoxyacetic acid [226] with HSA, green fluorescent fusion protein with cyclophilin [45], detection of hidden ligands that stabilize cholinesterase conformation [227], investigation of metal complexes with metallothionein in rat tissues [228], kinetic parameters of rhodanase [229], effect of oxidation of low-density lipoproteins on drug binding affinity [230], formation of a covalent conjugate between microcystin Lr and protein phosphatase 2a [231], identification of drug-binding sites in HSA [109] and modifying sites of mono-PEGylated calcitonins [232], and mapping of stereoselective recognition sites of human serum transferrin [233].…”