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|Title:||Systematic evolution of ligands by exponential enrichment: tissue SELEX|
|Abstract:||This invention discloses high-affinity oligonucleotide ligands to complex tissue targets, specifically nucleic acid ligands having the ability to bind to complex tissue targets, and the methods for obtaining such ligands. Tissue targets comprise cells, subcellular components, aggregates or cells, collections of cells, and higher ordered structures. Specifically, nucleic acid ligands to red blood cells ghosts, endothelia of the blood brain and CSF-blood barriers, glioblastomas, and lymphomas are described.|
|Inventor(s):||Heilig; Joseph S. (Boulder, CO), Gold; Larry (Boulder, CO)|
|Assignee:||NeXstar Pharmaceuticals, Inc. (Boulder, CO)|
|Filing Date:||Aug 05, 1997|
|Claims:||1. A method for purifying or identifying a transporter molecule of a blood brain barrier or cerebral spinal fluid (CSF)-blood barrier tissue comprising: |
a) identifying a nucleic acid ligand to an epitope of said transporter molecule by the method comprising:
i) preparing a candidate mixture of nucleic acids;
ii) contacting said candidate mixture of nucleic acids with a blood brain barrier endothelial cell or a CSF-blood barrier epithelial cell, wherein nucleic acids having an increased affinity to said epitope of said transporter molecule relative to the candidate mixture and capable of being transported across said barrier cell may be partitioned from the remainder of the candidate mixture;
iii) partitioning the increased affinity nucleic acids from the remainder of the candidate mixture; and
iv) amplifying the increased affinity nucleic acids to yield a mixture of nucleic acids enriched for nucleic acids with relatively higher affinity and specificity for binding to said epitope of said transporter molecule and capable of transport across said barrier cell, whereby a nucleic acid ligand to said epitope of said transporter molecule may be identified;
b) purifying said transporter molecule of said blood brain barrier or CSF-blood barrier tissue away from the remainder of said tissue on the basis of affinity between said epitope and said nucleic acid ligand; and
c) optionally identifying said transporter molecule.
2. The method of claim 1 wherein said transporter molecule is selected from the group consisting of a protein, a lipid and a carbohydrate.
3. A purified transporter molecule identified according to the method of claim 1.
4. The purified transporter molecule of claim 1 which is selected from the group consisting of a protein, a lipid and a carbohydrate.