High-affinity RNA aptamers to C-reactive protein (CRP)

Newly developed pre-elution methods for aptamer selection

N. Orito, S. Umekage, K. Sato, S. Kawauchi, H. Tanaka, E. Sakai, T. Tanaka, Yo Kikuchi

Research output: Contribution to journalArticle

Abstract

We have developed a modified SELEX (systematic evolution of ligands by exponential enrichment) method to obtain RNA aptamers with high affinity to C-reactive protein (CRP). CRP is a clinical biomarker present in plasma, the level of which increases in response to infections and noninfectious inflammation. The CRP level is also an important prognostic indicator in patients with several syndromes. At present, CRP content in blood is measured immunochemically using antibodies. To develop a more sensitive method using RNA aptamers, we have attempted to obtain high-affinity RNA aptamers to CRP. We succeeded in obtaining an RNA aptamer with high affinity to CRP using a CRP-immobilized Sepharose column and pre-elution procedure. Pre-elution is a method that removes the weak binding portion from a selected RNA population by washing for a short time with buffer containing CRP. By surface plasmon-resonance (SPR) analysis, the affinity constant of this aptamer for CRP was calculated to be K D = 2.25×10 -9 (M). The secondary structure, contact sites with CRP protein, and application of this aptamer will be described.

Original languageEnglish
Article number012042
JournalJournal of Physics: Conference Series
Volume352
Issue number1
DOIs
Publication statusPublished - 2012
Externally publishedYes

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elution
affinity
proteins
biomarkers
washing
infectious diseases
antibodies
surface plasmon resonance
blood
buffers

ASJC Scopus subject areas

  • Physics and Astronomy(all)

Cite this

High-affinity RNA aptamers to C-reactive protein (CRP) : Newly developed pre-elution methods for aptamer selection. / Orito, N.; Umekage, S.; Sato, K.; Kawauchi, S.; Tanaka, H.; Sakai, E.; Tanaka, T.; Kikuchi, Yo.

In: Journal of Physics: Conference Series, Vol. 352, No. 1, 012042, 2012.

Research output: Contribution to journalArticle

Orito, N. ; Umekage, S. ; Sato, K. ; Kawauchi, S. ; Tanaka, H. ; Sakai, E. ; Tanaka, T. ; Kikuchi, Yo. / High-affinity RNA aptamers to C-reactive protein (CRP) : Newly developed pre-elution methods for aptamer selection. In: Journal of Physics: Conference Series. 2012 ; Vol. 352, No. 1.
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