کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
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1174108 | 961729 | 2012 | 4 صفحه PDF | دانلود رایگان |
![عکس صفحه اول مقاله: Development of a fluorescence polarization binding assay for asialoglycoprotein receptor Development of a fluorescence polarization binding assay for asialoglycoprotein receptor](/preview/png/1174108.png)
Asialoglycoprotein receptor (ASGP-R) has been actively investigated for targeted delivery of therapeutic agents into hepatocytes because this receptor is selectively and highly expressed in liver and has a high internalization rate. Synthetic cluster glycopeptides (e.g., triGalNAc) bind with high affinity to ASGP-R and, when conjugated to a therapeutic agent, can drive receptor-mediated uptake in liver. We developed a novel fluorescent polarization (FP) ASGP-R binding assay to determine the binding affinities of ASGP-R-targeted molecules. The assay was performed in 96-well microplates using membrane preparations from rat liver as a source of ASGP-R and Cy5 fluorophore-labeled triGalNAc synthetic ligand as a tracer. This high-throughput homogeneous assay demonstrates advantages over existing multistep methods in that it minimizes both time and resources spent in determining binding affinities to ASGP-R. At the optimized conditions, a Z′ factor of 0.73 was achieved in a 96-well format.
Journal: Analytical Biochemistry - Volume 425, Issue 1, 1 June 2012, Pages 43–46