A two-dimensional suspension array system by coupling field flow fractionation to flow cytometry.
Flow field flow fractionation (Fl-FFF) was coupled to flow cytometry to improve the performance of suspension arrays. Size-based separation of the protein-conjugated microspheres by Fl-FFF was performed and the results demonstrated that, the separation could tolerate a wide range of carrier fluid conditions (pH values, salt concentrations, and buffer compositions) favorable for immunoassays. The immuno-complex remained intact during Fl-FFF, as revealed by fluorescence measurements before and after the Fl-FFF separation, and SDS-PAGE of the eluted proteins. The sample throughput of the suspension array can be increased several folds by using particles of different sizes and separating them with Fl-FFF before flow cytometric measurement. Moreover, the gel result hinted that the continuous wash inside the Fl-FFF system may lower the assay background, another possible advantage of the two-dimensional suspension array system.
Li
Jishan
J
Department of Chemistry, University of California, Riverside, CA 92521, USA.
Zhong
Wenwan
W
D000936:Antigen-Antibody Complex / Q000302:isolation & purification; D002021:Buffers; D005434:Flow Cytometry / Q000379:methods*; D031521:Fractionation, Field Flow / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D007118:Immunoassay / Q000379:methods; D008863:Microspheres
D000936:Antigen-Antibody Complex; D002021:Buffers