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-   -   [NMR paper] Cell-compatible, integrin-targeted cryptophane-(129)Xe NMR biosensors. (http://www.bionmr.com/forum/journal-club-9/cell-compatible-integrin-targeted-cryptophane-129-xe-nmr-biosensors-21444/)

nmrlearner 11-06-2014 08:45 PM

Cell-compatible, integrin-targeted cryptophane-(129)Xe NMR biosensors.
 
Cell-compatible, integrin-targeted cryptophane-(129)Xe NMR biosensors.

Related Articles Cell-compatible, integrin-targeted cryptophane-(129)Xe NMR biosensors.

Chem Sci. 2011 Jun;2(6):1103-1110

Authors: Seward GK, Bai Y, Khan NS, Dmochowski IJ

Abstract
Peptide-modified cryptophane enables sensitive detection of protein analytes using hyperpolarized (129)Xe NMR spectroscopy. Here we report improved targeting and delivery of cryptophane to cells expressing ?v?3 integrin receptor, which is overexpressed in many human cancers. Cryptophane was functionalized with cyclic RGDyK peptide and Alexa Fluor 488 dye, and cellular internalization was monitored by confocal laser scanning microscopy. Competitive blocking assays confirmed cryptophane endocytosis through an ?v?3 integrin receptor-mediated pathway. The peptide-cryptophane conjugate was determined to be nontoxic in normal human lung fibroblasts by MTT assay at the micromolar cryptophane concentrations typically used for hyperpolarized (129)Xe NMR biosensing experiments. Flow cytometry revealed 4-fold higher cellular internalization in cancer cells overexpressing the integrin receptor compared to normal cells. Nanomolar inhibitory concentrations (IC 50 = 20-30 nM) were measured for cryptophane biosensors against vitronectin binding to ?v?3 integrin and fibrinogen binding to ?IIb?3 integrin. Functionalization of the conjugate with two propionic acid groups improved water solubility for hyperpolarized (129)Xe NMR spectroscopic studies, which revealed a single resonance at 67 ppm for the (129)Xe-cryptophane-cyclic RGDyK biosensor. Introduction of ?IIb?3 integrin receptor in detergent solution generated a new "bound" (129)Xe biosensor peak that was shifted 4 ppm downfield from the "free" (129)Xe biosensor.


PMID: 25364495 [PubMed - as supplied by publisher]



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