Separation method based on affinity reaction between magnetic and nonmagnetic particles for the analysis of particles and biomolecules.
A separation method is reported for particle and biochemical analysis based on affinity interactions between particle surfaces under magnetic field. In this method, magnetic particles with immunoglobulin G (IgG) or streptavidin on the surface are flowed through a separation channel to form a deposition matrix for selectively capturing nonmagnetic analytes with protein A or biotin on the surface due to specific antigen (Ag)--antibody (Ab) interactions. This separation method was demonstrated using model reactions of IgG--protein A and streptavidin-biotin on particle surface. The features of this new separation method are (1) the deposited Ag-Ab complex can be examined and further analyzed under the microscope, (2) a kinetic study of complex binding is possible, and (3) the predeposited matrix can be formed selectively and changed easily. The detection limits were about 10(-11) g. The running time was less than 10 min. The selectivities of studied particles were 94% higher than those of label-controlled particles. This method extends the applications of analytical magnetapheresis to nonmagnetic particles. Preliminary study shows that this separation method has a great potential to provide a simple, fast, and selective analysis for particles, blood cells, and immunoassay related applications.
Tsai
H Y
HY
School of Applied Chemistry, Chung Shan Medical University, Taichung, Taiwan.
Hsu
F H
FH
Lin
Y P
YP
Bor Fuh
C
C
D001710:Biotin / Q000032:analysis / Q000378:metabolism; D002846:Chromatography, Affinity / Q000379:methods*; D007074:Immunoglobulin G / Q000032:analysis / Q000378:metabolism; D008280:Magnetics / Q000295:instrumentation*; D010316:Particle Size; D011485:Protein Binding; D015203:Reproducibility of Results; D013205:Staphylococcal Protein A / Q000032:analysis / Q000378:metabolism; D019809:Streptavidin / Q000032:analysis / Q000378:metabolism
D007074:Immunoglobulin G; D013205:Staphylococcal Protein A; D001710:Biotin; D019809:Streptavidin