Particle electrophoresis of micrometric-sized superparamagnetic particles designed for magnetic purification of cells.
Measuring the electrophoretic mobility of superparamagnetic particles (0.5-4.5 microns mean size) was undertaken to probe the coupling of lectins and antibodies to their surface. Coupling was either noncovalent (antigen-antibody and biotin-streptavidin linkage) or covalent (tosyl-activated beads). The direct observation of the electrophoresis of single particles illuminated in dark field and processed by image analysis allowed the determination of their apparent electrophoretic mobility. Mobilities ranged from -0.5 micron s-1/CmV-1 to +1 micron s-1/CmV-1 when measured at 20 degrees C in 0.15 M NaCl and 30 mg/mL sorbitol, pH 7.4. The relative standard deviation was less than 0.1%. Surface immobilization of charged proteins onto the superparamagnetic beads shifted their electrophoretic mobility up to 200%; this was also quantitatively correlated with some specific properties (enzymatic activity, antigen-binding activity, lectin-binding activity). Although particle electrophoresis has mainly been reported for the study of surface adsorption phenomenon, it may be a versatile tool for controlling covalent modifications of particles designed for therapeutical targeting or chromatographic use and may also apply to a quantitative analysis of ligand-binding interactions.
Sestier
C
C
INSERM Unit 313, Hopital Pitié Salpétrière, Paris, France. sestierclaude@minitel.net
Sabolovic
D
D
D001710:Biotin; D002460:Cell Line; D004586:Electrophoresis / Q000379:methods*; D004727:Endothelium / Q000166:cytology; D004730:Endothelium, Vascular / Q000166:cytology; D006801:Humans; D007075:Immunoglobulin M; D018189:Immunomagnetic Separation / Q000379:methods*; D037102:Lectins; D008863:Microspheres; D010316:Particle Size
D007075:Immunoglobulin M; D037102:Lectins; D001710:Biotin