Sedimentation field flow fractionation of immunoglobulin A coated polystyrene beads. Influence of carrier composition on complex characterization.
The amount of immunoglobulin A (IgA) adsorbed on the surface of two different samples of polystyrene (PS) microbeads was evaluated using differential sedimentation field flow fractionation (SdFFF) analyses. For the first time, the SdFFF separations obtained by using, as mobile phase, solutions common to many biochemical procedures and applications have been compared and discussed. Good separation results were achieved in the different carriers, and the SdFFF gave equivalent mass per particle values in all carriers provided that the pH and ionic strength conditions of the eluents were well controlled. The IgA adsorption process onto PS occurred by maintaining unaltered the capacity of the PS-IgA substrate to selectively recognize anti-IgA (aIgA), as proven by elution of the ternary complex PS-IgA-aIgA and from the monitored lack of reaction when the PS-IgA was placed in contact with aIgE.
Contado
Catia
C
Department of Chemistry, University of Ferrara, Via L. Borsari, 46, I-44100 Ferrara, Italy. catia.contado@unife.it
Bregola
Letizia
L
Dondi
Francesco
F
D000327:Adsorption; D000936:Antigen-Antibody Complex / Q000032:analysis / Q000302:isolation & purification*; D002021:Buffers; D002498:Centrifugation; D031521:Fractionation, Field Flow / Q000295:instrumentation* / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D007070:Immunoglobulin A / Q000032:analysis / Q000302:isolation & purification*; D007073:Immunoglobulin E / Q000302:isolation & purification; D008863:Microspheres; D008970:Molecular Weight; D009994:Osmolar Concentration; D010316:Particle Size; D011137:Polystyrenes / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D013056:Spectrophotometry, Ultraviolet / Q000379:methods; D013499:Surface Properties
D000936:Antigen-Antibody Complex; D002021:Buffers; D007070:Immunoglobulin A; D011137:Polystyrenes; D007073:Immunoglobulin E