Abstract：
:High affinity aptamer-based biomarker discovery has the advantage of simultaneously discovering an aptamer affinity reagent and its target biomarker protein. Here, we demonstrate a morphology-based tissue aptamer selection method that enables us to use tissue sections from individual patients and identify high-affinity aptamers and their associated target proteins in a systematic and accurate way. We created a combinatorial DNA aptamer library that has been modified with thiophosphate substitutions of the phosphate ester backbone at selected 5´dA positions for enhanced nuclease resistance and targeting. Based on morphological assessment, we used image-directed laser microdissection (LMD) to dissect regions of interest bound with the thioaptamer (TA) library and further identified target proteins for the selected TAs. We have successfully identified and characterized the lead candidate TA, V5, as a vimentin-specific sequence that has shown specific binding to tumor vasculature of human ovarian tissue and human microvascular endothelial cells. This new Morph-X-Select method allows us to select high-affinity aptamers and their associated target proteins in a specific and accurate way, and could be used for personalized biomarker discovery to improve medical decision-making and to facilitate the development of targeted therapies to achieve more favorable outcomes.
journal_name
Biotechniquesjournal_title
BioTechniquesauthors
Wang H,Li X,Volk DE,Lokesh GL,Elizondo-Riojas MA,Li L,Nick AM,Sood AK,Rosenblatt KP,Gorenstein DGdoi
10.2144/000114473subject
Has Abstractpub_date
2016-11-01 00:00:00pages
249-259issue
5eissn
0736-6205issn
1940-9818pii
000114473journal_volume
61pub_type
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