Heparin-immobilized microspheres for the capture of cytokines.
The preparation and characterization of heparin-immobilized microspheres which were used to bind acidic fibroblast growth factor (aFGF), vascular endothelial growth factor (VEGF), monocyte chemoattractant protein 1 (MCP-1/CCL2), and regulation upon activation normal T cell express sequence (RANTES/CCL5) is described. These beads were used as trapping agents in microdialysis sampling experiments in a separate study. Both free heparin and a synthesized heparin-albumin conjugate were immobilized onto microspheres and compared for their effectiveness. The heparin-albumin conjugate microspheres exhibited significant nonspecific adsorption which appeared to be due to the albumin content. The prepared heparin-immobilized microspheres were stable for 3 months at 4 °C. A bead-based flow cytometric assay was developed to study the binding capacity and specificity of the heparin-immobilized microspheres to cytokines. These heparin-immobilized microspheres exhibited broad dynamic ranges for binding to the four cytokines (aFGF, 1.0-1,000 ng/mL; VEGF, 0.5-1,000 ng/mL; CCL2, 1.95-1,000 ng/mL; CCL5, 1.95-500 ng/mL). Fast binding kinetics of the cytokines to the heparin-immobilized beads suggests that these beads may be useful as affinity agents in microfluidic flow systems.
Duo
Jia
J
Department of Chemistry and Chemical Biology, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY 12180, USA.
Stenken
Julie A
JA
D000818:Animals; D002417:Cattle; D018932:Chemokine CCL2 / Q000302:isolation & purification / Q000378:metabolism; D018946:Chemokine CCL5 / Q000302:isolation & purification / Q000378:metabolism; D016207:Cytokines / Q000302:isolation & purification* / Q000378:metabolism; D016220:Fibroblast Growth Factor 1 / Q000302:isolation & purification / Q000378:metabolism; D005434:Flow Cytometry / Q000379:methods; D006493:Heparin / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D055767:Immobilized Proteins / Q000737:chemistry / Q000378:metabolism; D008863:Microspheres; D011485:Protein Binding; D051381:Rats; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A / Q000302:isolation & purification / Q000378:metabolism
D018932:Chemokine CCL2; D018946:Chemokine CCL5; D016207:Cytokines; D055767:Immobilized Proteins; D042461:Vascular Endothelial Growth Factor A; D016220:Fibroblast Growth Factor 1; D012710:Serum Albumin, Bovine; D006493:Heparin
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