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Details for Patent: 5,712,375

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Details for Patent: 5,712,375

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, glioblastomas, and lymphomas are described.
Inventor(s): Jensen; Kirk B. (Boulder, CO), Chen; Hang (Boulder, CO), Morris; Kevin N. (Schworzach, AT), Stephens; Andrew (Denver, CO), Gold; Larry (Boulder, CO)
Assignee: NeXstar Pharmaceuticals, Inc. (Boulder, CO)
Filing Date:May 03, 1995
Application Number:08/434,001
Claims:1. A method for purifying and isolating a protein component of a biological tissue comprising:

a) identifying a nucleic acid ligand to a protein component of a biological tissue by the method comprising:

i) preparing a candidate mixture of nucleic acids;

ii) contacting said candidate mixture of nucleic acids with said biological tissue, wherein nucleic acids having an increased affinity to the biological tissue relative to the candidate mixture may be partitioned from the remainder of the candidate mixture and wherein said nucleic acids have a specific affinity to a protein component of said biological tissue;

iii) partitioning the increased affinity nucleic acids from the remainder of the candidate mixture;

iv) amplifying the increased affinity nucleic acids to yield a mixture of nucleic acids enriched for nucleic acids with an increased affinity and an increased specificity for binding to said protein component; and

v) identifying said nucleic acid ligand of said protein component; and

b) purifying and isolating said protein component of said biological tissue away from said biological tissue on the basis of affinity between said protein component and said nucleic acid ligand.

2. The method of claim 1 wherein said candidate mixture is comprised of single-stranded nucleic acids.

3. The method of claim 2 wherein the single-stranded nucleic acids are deoxyribonucleic acids.

4. The method of claim 2 wherein the single-stranded nucleic acids are ribonucleic acids.

5. The method of claim 1 wherein said biological tissue is selected from the group consisting of red blood cell ghosts, glioblastoma, and lymphoma.

6. A method for purifying and isolating a protein component of a biological tissue comprising:

a) identifying a nucleic acid ligand to a protein component of a biological tissue by the method comprising:

i) preparing a candidate mixture of nucleic acids;

ii) contacting the candidate mixture with a first biological tissue, wherein nucleic acids having an increased affinity to the first biological tissue relative to the candidate mixture may be partitioned from the remainder of the candidate mixture and wherein said nucleic acids have a specific affinity to a protein component of said first biological tissue;

iii) partitioning the increased affinity nucleic acids from the remainder of the candidate mixture;

iv) contacting the increased affinity nucleic acids with a second biological tissue, wherein nucleic acids with affinity to the second biological tissue are removed;

v) amplifying the remaining nucleic acids with specific affinity to said protein component to yield a mixture of nucleic acids enriched for nucleic acids with relatively higher affinity and specificity for binding to said protein component; and

vi) identifying said nucleic acid ligand of said protein component; and

b) purifying and isolating said protein component of said first biological tissue away from said first biological tissue on the basis of affinity between said protein Component and said nucleic acid ligand.

7. A method for purifying and isolating a protein component of a biological tissue comprising:

a) identifying a nucleic acid ligand to a protein component of a biological tissue by the method comprising:

i) preparing a candidate mixture of nucleic acids;

ii) contacting the candidate mixture with a first biological tissue, wherein nucleic acids with affinity to the first biological tissue are removed from the candidate mixture;

iii) contacting the candidate mixture from (ii) with a second biological tissue, wherein nucleic acids having an increased affinity to the second biological tissue relative to the candidate mixture may be partitioned from the remainder of the candidate mixture and wherein said nucleic acids have a specific affinity to a protein component of said second biological tissue;

iv) partitioning the increased affinity nucleic acids from the remainder of the candidate mixture;

v) amplifying the nucleic acids with specific affinity to said protein component to yield a mixture of nucleic acids enriched for nucleic acids with relatively higher affinity and specificity for binding to said protein component; and

vi) identifying the nucleic acid ligand of said protein; and

b) purifying and isolating said protein component of said second biological tissue away from said second biological tissue on the basis of affinity between said protein component and said nucleic acid ligand.
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