Aptamer-Modified Magnetic Beads in Affinity Separation of Proteins

Part of the Methods in Molecular Biology book series (MIMB, volume 1286)

Abstract

Aptamers are valuable alternative ligands for affinity separations. Here, we describe the aptamer-based affinity separation of His-tagged proteins using an aptamer directed against the His-tag. The immobilization of the aptamer to magnetic beads is described as well as the aptamer-based purification and proper methods for the characterization of the process. Moreover, indications for the transfer of the process to other aptamers are given.

Key words

Aptamer Affinity separation His-tag Protein purification Magnetic beads 

Notes

Acknowledgement

This work was supported by the German Research Foundation DFG.

References

  1. 1.
    Ellington AD, Szostak JW (1990) In vitro selection of RNA molecules that bind specific ligands. Nature 346:818–822CrossRefPubMedGoogle Scholar
  2. 2.
    Robertson DL, Joyce GF (1990) Selection in vitro of an RNA enzyme that specifically cleaves single-stranded DNA. Nature 344:467–468CrossRefPubMedGoogle Scholar
  3. 3.
    Tuerk C, Gold L (1990) Systematic evolution of ligands by exponential enrichment: RNA ligands to bacteriophage T4 DNA polymerase. Science 249:505–510CrossRefPubMedGoogle Scholar
  4. 4.
    Jayasena SD (1999) Aptamers: an emerging class of molecules that rival antibodies in diagnostics. Clin Chem 45:1628–1650PubMedGoogle Scholar
  5. 5.
    Walter J-G, Stahl F, Scheper T (2012) Aptamers as affinity ligands for downstream processing. Eng Life Sci 12:496–506CrossRefGoogle Scholar
  6. 6.
    Romig TS, Bell C, Drolet DW (1999) Aptamer affinity chromatography: combinatorial chemistry applied to protein purification. J Chromatogr B Biomed Sci Appl 731:275–284CrossRefPubMedGoogle Scholar
  7. 7.
    Doyle SA, Murphy MB (2008) Aptamers and methods for their in vitro selection and uses thereof. P.A. Publication. US 7329742(10/934,856)Google Scholar
  8. 8.
    Kökpinar Ö, Walter J-G, Shoham Y, Stahl F, Scheper T (2011) Aptamer-based downstream processing of His-tagged proteins utilizing magnetic beads. Biotechnol Bioeng 108:2371–2379CrossRefPubMedGoogle Scholar
  9. 9.
    Krebsfanger N, Schierholz K, Bornscheuer UT (1998) Enantioselectivity of a recombinant esterase from Pseudomonas fluorescens towards alcohols and carboxylic acids. J Biotechnol 60:105–111CrossRefPubMedGoogle Scholar
  10. 10.
    Meng J, Walter J, Koekpinar O, Stahl F, Scheper T (2008) Automated microscale His-tagged protein purification using Ni-NTA magnetic agarose beads. Chem Eng Technol 31:463–468CrossRefGoogle Scholar
  11. 11.
    Walter JG, Kokpinar O, Friehs K, Stahl F, Scheper T (2008) Systematic investigation of optimal aptamer immobilization for protein-microarray applications. Anal Chem 80:7372–7378CrossRefPubMedGoogle Scholar
  12. 12.
    Zhu G, Lübbecke M, Walter J, Stahl F, Scheper T (2011) Characterization of optimal aptamer-microarray binding chemistry and spacer design. Chem Eng Technol 34:2022–2028CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.Institut für Technische ChemieGottfried Wilhelm Leibniz Universität HannoverHannoverGermany

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