• Bulletin of the Korean Chemical Society
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• 제35권9호
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• pp.2665-2668
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• 2014

Aptamers are oligonucleotides or peptide molecules that are able to bind to their specific target molecules with high affinity via molecular recognition. In this study, we present development of aptamer-based precipitation assays (or simply aptamoprecipitation) for His-tagged proteins and thrombin to compare their purification efficiency with other conventional affinity precipitation methods. A crosslinking method was employed to immobilize thiol-functionalized aptamers onto the surface of polystyrene resins, enabling them to specifically bind to His-tag and to thrombin, respectively. The resulting aptamer-functionalized resins were successfully applied via a one-step experiment to purification of His-tagged proteins from complex E. coli and to thrombin extraction, exhibiting superior or at least comparable purification results to the conventional immobilized metal affinity precipitation or immunoprecipitation.

• Mass Spectrometry Letters
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• 제3권3호
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• pp.82-85
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• 2012

The formation of zinc finger peptide-$Zn^{2+}$ complexes in electrospray ionization mass spectrometry (ESI-MS) was examined using three different metal ion sources: $ZnCl_2$, $Zn(CH_3COO)_2$, and $Zn(OOC(CHOH)_2COO)$. For the four zinc finger peptides (Sp1-1, Sp1-3, CF2II-4, and CF2II-6) that bind only a single $Zn^{2+}$ in the native condition, electrospray of apo-zinc finger in solution containing $ZnCl_2$ or $Zn(CH_3COO)_2$ resulted in the formation of zinc finger-$Zn^{2+}$ complexes with multiple zinc ions. This result suggests the formation of nonspecific zinc finger-$Zn^{2+}$ complexes. Zn(tartrate), $Zn(OOC(CHOH)_2COO)$, mainly produced specific zinc finger-$Zn^{2+}$ complexes with a single zinc ion. This study clearly indicates that tartrate is an excellent counter ion in ESI-MS studies of zinc finger-$Zn^{2+}$ complexes, which prevents the formation of nonspecific zinc finger-$Zn^{2+}$ complexes.