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In vitro and in vivo affinity microdialysis sampling of cytokines using heparin-immobilized microspheres

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Abstract

Heparin-immobilized microspheres were included in microdialysis sampling perfusion fluids under both in vitro and in vivo conditions to improve the recovery of different cytokines, acidic fibroblast growth factor, vascular endothelial growth factor, monocyte chemoattractant protein-1 (or CCL2), and regulation upon activation normal T cell express sequence (or CCL5). Different strategies to dissociate captured CCL2 and CCL5 from the immobilized heparin were attempted, and both cytokines could be quantitatively eluted from the beads using a phosphate buffer (pH 7.4) containing 25% (v/v) acetonitrile which did not interfere with the subsequent detection of cytokine using an ELISA assay. Using these heparin-immobilized microspheres, a two to fivefold increase of microdialysis relative recovery (RR) was achieved for the four cytokines from a quiescent solution. Enhanced microdialysis RR of CCL2 using the heparin-immobilized microspheres from microdialysis probes implanted into the peritoneal cavity of a rat was performed to test the in vivo application. This work suggests that the heparin-immobilized microspheres provide an alternative affinity agent to the previously used antibody-immobilized microspheres for enhanced microdialysis sampling of cytokines.

Microdialysis sampling with heparin-immobilized microspheres has been used to capture cytokines aFGF, CCL2, CCL5, and VEGF

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Acknowledgment

We thank NIH EB 001441 for the funding support of this work. Part of this work was also supported by the Arkansas Biosciences Institute, the major research component of the Arkansas Tobacco Settlement Proceeds Act of 2000. Professor Robert J. Linhardt, Rensselaer Polytechnic Institute, is gratefully acknowledged for the discussion of heparin/cytokine interactions. We also thank Dr. Fuming Zhang (Rensselaer Polytechnic Institute) for the assistance with heparin immobilization chemistry and Steven Lotz and Yili Lin (Albany Medical College) for the assistance with flow cytometric analysis.

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  1. Jia Duo

    Present address: Forchheimer G19, Albert Einstein College of Medicine of Yeshiva University, 1300 Morris Park Avenue, Bronx, NY, 10461, USA

Authors and Affiliations

  1. Department of Chemistry and Chemical Biology, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY, 12180, USA

    Jia Duo

  2. Department of Chemistry and Biochemistry, University of Arkansas, Fayetteville, AR, 72701, USA

    Julie A. Stenken

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  1. Jia Duo
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  2. Julie A. Stenken
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Correspondence to Julie A. Stenken.

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Published in the special issue Heparin Characterization with Guest Editor Cynthia K. Larive.

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Duo, J., Stenken, J.A. In vitro and in vivo affinity microdialysis sampling of cytokines using heparin-immobilized microspheres. Anal Bioanal Chem 399, 783–793 (2011). https://doi.org/10.1007/s00216-010-4333-0

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  • DOI: https://doi.org/10.1007/s00216-010-4333-0

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