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A two-dimensional suspension array system by coupling field flow fractionation to flow cytometry

Authors
Journal
Journal of Chromatography A
0021-9673
Publisher
Elsevier
Publication Date
Volume
1183
Identifiers
DOI: 10.1016/j.chroma.2008.01.029
Keywords
  • Flow Field Flow Fractionation
  • Size Separation
  • Suspension Array
  • Immunoassay
  • Array Throughput
Disciplines
  • Biology

Abstract

Abstract 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.

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