Biotechnology Letters

, Volume 32, Issue 3, pp 379–385 | Cite as

Selection of RNA aptamers specific to active prostate-specific antigen

  • Sujin Jeong
  • Seung Ryul Han
  • Young Ju Lee
  • Seong-Wook Lee
Original Research Paper


A counter-SELEX procedure with recombinant purified active prostate specific antigen (PSA) was used to identify specific RNA aptamers against the active PSA. We developed two different kinds of counter-SELEX methods; one includes pre-clearance step with inactive proPSA protein, and the other with tagged GST protein. After 9 iterative selection cycles, several identical RNA aptamers can be identified from both counter-SELEX methods. Real-time PCR analysis and gel retardation experiment showed that the aptamers have a specific binding activity against the active PSA, but not for GST or proPSA. These aptamers could be of potential use as specific diagnostic, imaging and/or therapeutic agents against prostate cancer.


Prostate cancer Prostate-specific antigen RNA aptamer SELEX 



This subject is supported by Korea Ministry of Environment as “The Eco-technopia 21 project” (091-081-073) and Bio Strategic Technology Development of Korea Ministry of Knowledge Economy (10031930).


  1. Balk SP, Ko YJ, Bubley GJ (2003) Biology of prostate-specific antigen. J Clin Oncol 21:383–391CrossRefPubMedGoogle Scholar
  2. Brawer MK (1999) Prostate-specific antigen: current status. CA Cancer J Clin 49:264–281CrossRefPubMedGoogle Scholar
  3. Bunka DH, Stockley PG (2006) Aptamers come of age-at last. Nat Rev Microbiol 4:588–596CrossRefPubMedGoogle Scholar
  4. Catalona WJ, Smith DS, Ratliff TL, Dodds K, Coplen DE, Yuan JJ, Petros JA, Andriole GL (1991) Measurement of prostate-specific antigen in serum as a screening test for prostate cancer. N Engl J Med 324:1156–1161PubMedCrossRefGoogle Scholar
  5. Hwang B, Cho JS, Yeo HJ, Kim JH, Chung KM, Han K, Jang SK, Lee SW (2004) Isolation of specific and high-affinity RNA aptamers against NS3 helicase domain of hepatitis C virus. RNA 10:1277–1290CrossRefPubMedGoogle Scholar
  6. Hwang B, Lim JH, Hahm B, Jang S, Lee SW (2009) hnRNP L is required for the translation mediated by HCV IRES. Biochem Biophys Res Commun 378:584–588CrossRefPubMedGoogle Scholar
  7. Jeong S, Lee SW (2007) Expression and purification of recombinant active prostate-specific antigen from Escherichia coli. J Microbiol Biotechnol 17:840–846PubMedGoogle Scholar
  8. Kumar A, Mikolajczyk SD, Goel AS, Millar LS, Saedi MS (1997) Expression of pro form of prostate-specific antigen by mammalian cells and its conversion to mature, active form by human kallikrein 2. Cancer Res 57:3111–3114PubMedGoogle Scholar
  9. LeBeau AM, Singh P, Isaacs JT, Denmeade SR (2008) Potent and selective peptidyl boronic acid inhibitors of the serine protease prostate-specific antigen. Chem Biol 15:665–674CrossRefPubMedGoogle Scholar
  10. Michel S, Collomb-Clere E, Geourjon C, Charrier J, Passagot J, Courty Y, Deleage G, Jolivet-Reynaud C (2005) Selective recognition of enzymatically active prostate specific antigen (PSA) by anti-PSA monoclonal antibodies. J Mol Recognit 18:225–235CrossRefPubMedGoogle Scholar
  11. Que-Gewirth NS, Sullenger BA (2007) Gene therapy progress and prospects: RNA aptamers. Gene Ther 14:283–291CrossRefPubMedGoogle Scholar
  12. Stenman UH, Leinonen J, Alfthan H, Rannikko S, Tuhkanen K, Alfthan O (1991) A complex between prostate-specific antigen and alpha 1-antichymotrypsin is the major form of prostate-specific antigen in serum of patients with prostatic cancer: assay of the complex improves clinical sensitivity for cancer. Cancer Res 51:222–226PubMedGoogle Scholar
  13. Sutkowski DM, Goode RL, Baniel J, Teater C, Cohen PA, McNulty M, Hsiung HM, Becker GW, Neubauer BL (1999) Growth regulation of prostatic stromal cells by prostate-specific antigen. J Natl Cancer Inst 91:1663–1669CrossRefPubMedGoogle Scholar
  14. Takayama TK, Fujikawa K, Davie EW (1997) Characterization of the precursor of prostate specific antigen: activation by trypsin and by human glandular kallikrein. J Biol Chem 272:21582–21588CrossRefPubMedGoogle Scholar
  15. Webber MM, Waghray A, Bello D (1995) Prostate-specific antigen, a serine protease, facilitates human prostate cancer invasion. Clin Cancer Res 1:1089–1094PubMedGoogle Scholar
  16. Wu P, Stenman U, Pakkala M, Narvanen A, Leinonen J (2004) Separation of enzymatically active and inactive prostate-specific antigen (PSA) by peptide affinity chromatography. Prostate 58:345–353CrossRefPubMedGoogle Scholar
  17. Zuker M (2003) Mfold web server for nucleic acid folding and hybridization prediction. Nucleic Acids Res 31:3406–3415CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  • Sujin Jeong
    • 1
  • Seung Ryul Han
    • 1
  • Young Ju Lee
    • 1
  • Seong-Wook Lee
    • 1
  1. 1.Department of Molecular BiologyInstitute of Nanosensor and Biotechnology, Dankook UniversitySuji-gu, YonginKorea

Personalised recommendations