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Modified DNA Aptamer Immobilization via Cu(I)‐Stabilizing Ligand‐assisted Azide–Alkyne Cycloaddition for Surface Plasmon Resonance Measurement
Author(s) -
Kim NakHyeon,
Le Hoa Thi,
Yang YongSuk,
Byun Kyung Min,
Kim Tae Woo
Publication year - 2015
Publication title -
bulletin of the korean chemical society
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.237
H-Index - 59
ISSN - 1229-5949
DOI - 10.1002/bkcs.10524
Subject(s) - surface plasmon resonance , chemistry , cycloaddition , aptamer , azide , alkyne , click chemistry , electrospray ionization , amine gas treating , combinatorial chemistry , deoxyuridine , ligand (biochemistry) , surface modification , photochemistry , mass spectrometry , dna , nanotechnology , materials science , organic chemistry , nanoparticle , catalysis , biochemistry , receptor , chromatography , biology , genetics
The synthesis and binding integrity of an ErbB2-binding modified DNA aptamer (E2Ap) with 5-naphtyl-2′-deoxyuridine is confirmed by electrospray ionization mass spectrometry and by a 32P dot blotting experiment. E2Ap degradation in a conventional Cu(I)-catalyzed azide–alkyne cycloaddition (CuAAC) condition is efficiently prevented by using tris(3-hydroxypropyltriazolylmethyl)amine (THPTA). The immobilization of E2Ap on a surface plasmon resonance (SPR) chip is performed by azido-functionalization and THPTA-assisted CuAAC reaction. The folding, target binding, regeneration, and nonspecific interaction of E2Ap on the SPR chip are verified by both classical and kinetic titration experiments.