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Development of an optical RNA-based aptasensor for C-reactive protein

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Abstract

The development of a RNA-aptamer-based optical biosensor (aptasensor) for C-reactive protein (CRP) is reported. CRP is an important clinical biomarker; it was the first acute-phase protein to be discovered (1930) and is a sensitive systemic marker of inflammation and tissue damage. It has also a prognostic value for patients with acute coronary syndrome. The average concentration of CRP in serum is 0.8 ppm and it increases in response to a variety of inflammatory stimuli, such as trauma, tissue necrosis, infection and myocardial infarction. The interaction between the 44-base RNA aptamer and the target analyte CRP is studied. In particular, the influence of the aptamer immobilization procedure (chemistry, length, concentration), as well as the binding conditions, i.e., the influence on the binding of different buffers, the presence of Ca2+ ion and the specificity (against human serum albumin) have been evaluated. Using the best working conditions, we achieved a detection limit of 0.005 ppm, with good selectivity towards human serum albumin. Some preliminary experiments in serum are reported.

The assay on the CM5 chip

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References

  1. Jayasena SD (1999) Clin Chem 45:1628–1650

    CAS  Google Scholar 

  2. Tuerk C, Gold L (1990) Science 249:505–510

    Article  CAS  Google Scholar 

  3. Ellington AD, Szostak JW (1990) Nature 346:818–822

    Article  CAS  Google Scholar 

  4. Luzi E, Minunni M, Tombelli S, Mascini M (2003) Trends Anal Chem 22:810–818

    Article  CAS  Google Scholar 

  5. Tombelli, S, Minunni M, Mascini M (2005) Biosens Bioelectron 20:2424–2434

    Article  CAS  Google Scholar 

  6. Yang L, Fung CW, Cho EJ, Ellington AD (2007) Anal Chem 79:3320–3329

    Article  CAS  Google Scholar 

  7. Tombelli S, Minunni M, Mascini M (2007) Biomed Eng 24:191–200

    CAS  Google Scholar 

  8. Li YY, Zhang C, Li BS, Zhao LF, Li XB, Yang WJ, Xu SQ (2007) Clin Chem 56:1061–1066

    Article  CAS  Google Scholar 

  9. Tombelli S, Minunni M, Mascini M (2007) In: Van Emon JM (Ed) Immunoassays and other bioanalytical techniques. CRC, New York

    Google Scholar 

  10. Wang J (2007) Electroanalysis 19:769–776

    Article  CAS  Google Scholar 

  11. Hamula CLA, Guthrie JW, Zhang H, Li XF, Le XC (2006) Trends Anal Chem 25:681–691

    Article  CAS  Google Scholar 

  12. Lin C, Katilius E, Liu Y, Zhang J, Yan H (2006) Angew Chem Int Ed 45:5296–5301

    Article  CAS  Google Scholar 

  13. Maehashi K, Katsura T, Kerman K, Takamura Y, Matsumoto K, Tamiya E (2007) Anal Chem 79:782–787

    Article  CAS  Google Scholar 

  14. Li Y, Hye Jin Lee HJ, Corn RM (2007) Anal Chem 79:1082–1088

    Article  CAS  Google Scholar 

  15. Centi S, Tombelli S, Minunni M, Mascini M (2007) Anal Chem 79:1466–1473

    Article  CAS  Google Scholar 

  16. Bini A, Minunni M, Tombelli S, Centi S, Mascini M (2007) Anal Chem 79:3016–3019

    Article  CAS  Google Scholar 

  17. Xiao Y, Lubin AA, Heeger AJ, Plaxco KW (2005) Angew Chem Int Ed 44:5456–5459

    Article  CAS  Google Scholar 

  18. Heyduk E, Heyduk T (2005) Anal Chem 77:1147–1156

    Article  CAS  Google Scholar 

  19. Lai RY, Plaxco KW, Heeger AJ (2007) Anal Chem 79:229–233

    Article  CAS  Google Scholar 

  20. Clearfield MB (2005) J Am Osteopath Assoc 105:409–416

    Google Scholar 

  21. Ledue TB, Rifai N (2001) Clin Chem Lab Med 39:1171–1176

    Article  CAS  Google Scholar 

  22. Zhan W, Bar AJ (2007) Anal Chem 79:459–463

    Article  CAS  Google Scholar 

  23. Kartalov EP, Zhong JF, Scherer A, Quake SR, Taylor CR, Anderson WF (2006) BioTechniques 40:85–90

    CAS  Google Scholar 

  24. Kriz K, Ibraimi F, Lu M, Hansson LO, Kriz D (2005) Anal Chem 77:5920–5924

    Article  CAS  Google Scholar 

  25. Meyer MHF, Hartmann M, Krause HJ, Blankenstein G, Mueller-Chorus B, Oster J, Miethe P, Keusgen M (2007) Biosens Bioelectron 22:973–979

    Article  CAS  Google Scholar 

  26. Hu WP, Hsu HY, Chiou A, Tseng KY, Lin HY, Chang GL, Chen SJ (2006) Biosens Bioelectron 21:1631–1637

    Article  CAS  Google Scholar 

  27. Meyer MHF, Hartmann M, Keusgen M (2006) Biosens Bioelectron 21:1987–1990

    Article  CAS  Google Scholar 

  28. Lin S, Lee C-K, Lin Y-H, Lee S-Y, Sheu B-C, Tsai J-C, Hsu S-M, (2006) Biosens Bioelectron 22:715–721

    Article  CAS  Google Scholar 

  29. Jung S-H, Jung J-W, Suh I-B, Yuk JS, Kim W-J, Choi EY, Kim Y-M, Ha K-S (2007) Anal Chem 79:5703–5710

    Article  CAS  Google Scholar 

  30. Kim SD, Ryu JS, Yi HK, Kim SC, Zhang BT (2004) In: Proceedings of the 10th international meeting on DNA computing, pp 334–343

  31. Davis TM, Wilson WD (2001) Methods Enzymol 340:22–50

    Article  CAS  Google Scholar 

  32. Lofas S, Johnsson B (1990) J Chem Soc Chem Commun 21:1526–1528

    Article  Google Scholar 

  33. Ducongè F, Di Primo C, Toulmè JJ (2000) J Biol Chem 275:21287–21294

    Article  Google Scholar 

  34. Minunni M, Tombelli S, Gullotto A, Luzi E, Mascini M (2004) Biosens Bioelectron 20:1149–1156

    CAS  Google Scholar 

  35. Tombelli S, Minunni M, Luzi E, Mascini M (2005) Bioelectrochemistry 67:135–141

    Article  CAS  Google Scholar 

  36. Shrive AK, Gheetham GMT, Holden D, Myles DAA, Turnell WG, Volanakis JE, Pepys MB, Bloomer AC, Greenhough, TJ (1996) Nat Struct Mol Biol 3:346–354

    Article  CAS  Google Scholar 

  37. Potempa LA, Siegel JN, Fedel BA, Potempa RT, Gewurz H (1987) Mol Immunol 24:531–541

    Article  CAS  Google Scholar 

  38. Oliveira EB, Gotschlich EC, Liu TY (1977) Proc Natl Acad Sci USA 74:3148–3151

    Article  CAS  Google Scholar 

  39. Wang HW, Wu Y, Chen Y, Sui SF (2002) Int J Mol Med 9:665–671

    CAS  Google Scholar 

  40. Khreiss T, Jozsef L, Potempa A, Filep JG (2004) Circulation 109:2016–2022

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank the European Community for partially funding this work with the Integrated Project CARE-MAN, Healthcare by Biosensor Measurements and Networking (NMP4-CT-2006-017333).

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Correspondence to M. Mascini.

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Bini, A., Centi, S., Tombelli, S. et al. Development of an optical RNA-based aptasensor for C-reactive protein. Anal Bioanal Chem 390, 1077–1086 (2008). https://doi.org/10.1007/s00216-007-1736-7

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  • DOI: https://doi.org/10.1007/s00216-007-1736-7

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