Antibody-enhanced microdialysis collection of CCL2 from rat brain

by Herbaugh, Anthony W.; Stenken, Julie Ann

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Chemokine(C-C motif) Ligand 2 (CCL2 or MCP-1) is a signaling protein that is released under various conditions. In this study we demonstrate the first microdialysis collection of CCL2 from rat brain tissue using antibody-enhanced microdialysis. A monoclonal antibody to CCL2 was included in the dialysis perfusion fluid as an affinity agent to enhance the recovery of CCL2 both in vitro and in vivo. In vitro it was found that the use of antibody affinity agent increases the relative recovery of CCL2 from 9.6 ± 3.4% to 37.5 ± 10.2% and 64.8 ± 11.7% (n = 10) at flow rates of 2 μL/min and 1 μL/min, resp. Following the in vitro observation, CCL2 was collected from rat brain with microdialysis sampling using both control and antibody-included perfusion fluids. The in vivo data showed that relative recovery was increased at all but the first time point. This shows that the use of free antibody in the perfusion fluid increases the relative recovery of CCL2 and this enhanced microdialysis method may be applicable to other cytokines.

Journal
Journal of Neuroscience Methods
Volume
202
Issue
2
Year
2011
Start Page
124
ISBN/ISSN
0165-0270
PMID
21600925
DOI
10.1016/j.jneumeth.2011.05.006