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Session 5.3: Levy

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Albert Einstein Cancer Center
Albert Einstein Cancer Center
Health & MedicineTechnologyBusiness
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Levy Lab Matthew Levy Department of Biochemistry Albert Einstein College of Medicine Targeting cells with aptamers
Aptamers ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Aptamers exist naturally ,[object Object]
In vitro selection of aptamers Constant regions Random region Of 40 nucleotides Library size = ~10 14-15
Aptamers that target cell surface receptors can be used for delivery of cargoes to cells ,[object Object],[object Object],[object Object],Hicke et al. J Nucl Med. 2006 Apr;47(4):668-78 Anti-PSMA aptamer Anti-hTfR aptamer Anti-Tenascin C aptamer Aptamers can be used to target in vivo
Cancer/Aptamer projects underway in my lab ,[object Object],[object Object],[object Object],[object Object],[object Object]
The transferrin receptor (TfR, CD71) ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Selection for aptamers targeting the transferrin receptor ,[object Object],[object Object]
Anti-TfR aptamers label multiple human cancer cell lines FACs based analysis using AF488 labeled aptamer
Minimization eliminates more than half of the c2 sequence. c2 c2.min.2 c2.min.6 c2.min.9.1 38.6 kDa 14.0 kDa Brian Wengerter
Prostate-specific membrane antigen ,[object Object],[object Object],[object Object],[object Object],[object Object],Using a library based on the A9 aptamer we have now performed a ‘doped’ selection Cytoplasmic domain Transmembrane domain Extracellular domain NH 2 COOH Catalytic domain NH 2 COOH Catalytic domain
Doped A9 selection GGGAGGACGATGCGGACCGAAAAAGACCTGACTTCTATACTAAGTCTACGTTCCCAGACGACTCGCCC (68) Linsley Kelly Heavily mutagenize (30%) Reselect (HeLa-PSMA cells)
Minimization of the anti-PSMA A9 aptamer GGGAGGACGATGCGGACCGAAAAAGACCTGACTTCTATACTAAGTCTACGTTCCCAGACGACTCGCCC (68) Linsley Kelly
[object Object]
Targeting DEC 205 ,[object Object],[object Object],[object Object],[object Object]
Targeting cell surface receptors with aptamers anti-DEC205 ,[object Object],[object Object]
[object Object],Brian Wengerter Anti-DEC205 aptamers bind CHO cells that express DEC205
[object Object],[object Object],Brian Wengerter
Summary ,[object Object],[object Object],[object Object],[object Object]
Acknowledgements ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]

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Session 5.3: Levy

  • 1. Levy Lab Matthew Levy Department of Biochemistry Albert Einstein College of Medicine Targeting cells with aptamers
  • 2.
  • 3.
  • 4. In vitro selection of aptamers Constant regions Random region Of 40 nucleotides Library size = ~10 14-15
  • 5.
  • 6.
  • 7.
  • 8.
  • 9. Anti-TfR aptamers label multiple human cancer cell lines FACs based analysis using AF488 labeled aptamer
  • 10. Minimization eliminates more than half of the c2 sequence. c2 c2.min.2 c2.min.6 c2.min.9.1 38.6 kDa 14.0 kDa Brian Wengerter
  • 11.
  • 12. Doped A9 selection GGGAGGACGATGCGGACCGAAAAAGACCTGACTTCTATACTAAGTCTACGTTCCCAGACGACTCGCCC (68) Linsley Kelly Heavily mutagenize (30%) Reselect (HeLa-PSMA cells)
  • 13. Minimization of the anti-PSMA A9 aptamer GGGAGGACGATGCGGACCGAAAAAGACCTGACTTCTATACTAAGTCTACGTTCCCAGACGACTCGCCC (68) Linsley Kelly
  • 14.
  • 15.
  • 16.
  • 17.
  • 18.
  • 19.
  • 20.

Editor's Notes

  1. 20070212 combination of all data
  2. 20070212 combination of all data