Department of Chemistry and Biochemistry, University of Arkansas, Fayetteville, AR 72701, USA.
J Neurosci Methods. 2011 Nov 15;202(2):124-7. doi: 10.1016/j.jneumeth.2011.05.006. Epub 2011 May 11.
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 2μL/min and 1μL/min, respectively. 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.
趋化因子(C-C 基序)配体 2(CCL2 或 MCP-1)是一种在各种条件下释放的信号蛋白。在这项研究中，我们使用抗体增强型微透析首次从大鼠脑组织中采集 CCL2。将一种针对 CCL2 的单克隆抗体作为亲和剂包含在透析灌注液中，以增强 CCL2 在体外和体内的回收。体外研究发现，使用抗体亲和剂分别将流速为 2μL/min 和 1μL/min 时 CCL2 的相对回收率从 9.6±3.4%提高到 37.5±10.2%和 64.8±11.7%(n=10)。在体外观察之后，使用包含对照和抗体的灌注液通过微透析采样从大鼠脑组织中采集 CCL2。体内数据表明，除第一个时间点外，所有时间点的相对回收率均增加。这表明在灌注液中使用游离抗体可增加 CCL2 的相对回收率，并且这种增强型微透析方法可能适用于其他细胞因子。
