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Investigating Cytokine Binding Using a Previously Reported TNF-Specific Aptamer

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DOI: 10.4236/ojapps.2012.23019    4,812 Downloads   7,784 Views   Citations

ABSTRACT

Cytokines are of chief importance in the pathophysiology of sepsis and other systemic inflammatory response syndromes. We are designing and testing an extracorporeal cytokine adsorption device (CAD) that can remove cytokines via adsorption on biocompatible, microporous beads. The goal of this study was to determine whether a previously reported TNF binding DNA aptamer, 5’-GCGGCCGATA AGGTCTTTCC AAGCGAACGA ATTGAACCGC-3’, could be immobilized our hemoadsorption polymer surface to increase the removal rate of TNF. A reservoir consisting of horse serum spiked with a known concentration of TNF was perfused through our CAD packed with aptamer modified or unmodified (control) polymer beads. The binding affinity of the TNF aptamer was characterized using an enzyme-linked oligonucleotide assay (ELONA). As a positive control a well-established DNA aptamer that binds PDGF BB was also subjected to the same ELONA to validate the assay. TNF capture using the CAD showed no TNF removal over four hours for both the aptamer modified and unmodified control beads. Additionally, the results of the ELONA showed no binding of TNF to the reported aptamer; however the PDGF BB aptamer did bind PDGF BB. Based on these results we are able to conclude that the reported TNF specific aptamer does not bind TNF. These results will be of importance to other studies exploring aptamers for specific binding of TNF.

Conflicts of Interest

The authors declare no conflicts of interest.

Cite this paper

J. Fisher, M. DiLeo and W. Federspiel, "Investigating Cytokine Binding Using a Previously Reported TNF-Specific Aptamer," Open Journal of Applied Sciences, Vol. 2 No. 3, 2012, pp. 135-138. doi: 10.4236/ojapps.2012.23019.

References

[1] M. Schoenberg, M. Weiss, P. Radermacher, Outcome of patients with sepsis and septic shock after ICU treatment, Langenbeck's Arch Surg 383(1998) 44-48. doi:10.1007/s004230050090
[2] J.A. Kellum, L. Kong, M.P. Fink, L.A. Weissfield, D.M. Yealy, M.R. Pinsky, J. Fine, A. Krichevsky, R.L. Delude, D.C. Angus, Understanding the Inflammatory Cytokine Response in pneumonia and sepsis, Arch Intern Med 167(2007) 1655-1663. doi:10.1001/archinte.167.15.1655
[3] J. Kellum, Immunomodulation in sepsis: the role of hemofiltration, Minerva Anestesiol 65(1999) 410-418.
[4] J.A. Kellum, M.K. Dishart, Effect of hemofiltration filter adsorption on circulating IL-6 levels in septic rats, Crit Care 6(2002) 429-433. doi:10.1186/cc1528
[5] M. Song, J. Winchester, R.L. Albright, V.J. Capponi, M.D. Choquette, J.A. Kellum, Cytokine removal with a novel adsorbent polymer, Blood Purif 22(2004) 428-434. doi:10.1159/000080235
[6] M.V. DiLeo, J. Kellum, W.J. Federspiel, A simple mathematical model of cytokine capture using a hemoadsorption device, Annals of Biomedical Engineering 37(2008) 222-229. doi:10.1007/s10439-008-9587-8
[7] R. Stoltenburg, C. Reinemann, B. Strehlitz, SELEX--A (r)evolutionary method to generate high-affinity nucleic acid ligands, Biomolecular Engineering 24(2007) 381-403. doi:10.1016/j.bioeng.2007.06.001
[8] S.J. Klug, M. Famulok, All you wanted to know about SELEX, Molecular Biology Reports 20(1994) 97-107. doi:10.1007/BF00996358
[9] X. Yang, D.G. Gorenstein, Progress in thioaptamer development, Current Drug Targets 5(2004) 705-715. doi:10.2174/1389450043345074
[10] J.W. Guthrie, C.L.A. Hamula, H. Zhang, X.C. Le, Assays for cytokines using aptamers, Methods 38(2006) 324-330. doi:10.1016/j.ymeth.2006.01.001
[11] Z. Zhang, X. Yan, C. Xu, Oligonuecleotide antagonist for human tumor necrosis factor a (TNF-a), Institute for Viral Disease Control and Prevention Chinese Center for Disease Control and Prevention, United States, 2007.
[12] X. Yan, X. Gao, Z. Zhang, Isolation and Characterization of 2'-amino-modified RNA Aptamers for Human TNFa, Geno Prot Bioinfo 2(2004) 32-42.
[13] N. Zammatteo, C. Girardeaux, D. Delforge, J.-J. Pireaux, J. Remacle, Amination of polystyrene microwells: Application to the covalent grafting of DNA probes for hybridization assays, Analytical Biochemistry 236(1996) 85-94. doi:10.1006/abio.1996.0135
[14] M. Matteucci, M. Caruthers, Synthesis of deoxyoligonucleotides on a polymer support, J Am Chem Soc 103(1981) 3185-3191. doi:10.1021/ja00401a041
[15] L.S. Green, D. Jellinek, R. Jenison, A. Ostman, C.-H. Heldin, N. Janjic, Inhibitory DNA ligands to platelet-derived growth factor B-chain, Biochemistry 35(1996) 14413-14424. doi:10.1021/bi961544+

  
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