Modified DNA Aptamer Immobilization via Cu(I)-Stabilizing Ligand-assisted Azide-Alkyne Cycloaddition for Surface Plasmon Resonance Measurement
نویسندگان
چکیده
The synthesis and binding integrity of an ErbB2-binding modified DNA aptamer (E2Ap) with 5-naphtyl-20deoxyuridine is confirmed by electrospray ionization mass spectrometry and by a P 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 THPTAassisted 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.
منابع مشابه
Site-specific peptide and protein immobilization on surface plasmon resonance chips via strain-promoted cycloaddition.
Surface plasmon resonance (SPR) is a powerful label-free diagnostic tool to study biomolecular interactions. However, one of the drawbacks of SPR is the lack of controlled immobilization of ligands on the sensor surface. We have developed a modular platform for the fast, reagent-free and site-specific immobilization of azide-containing ligands by strain-promoted cycloaddition onto a cyclooctyne...
متن کاملCovalent attachment of diphosphine ligands to glassy carbon electrodes via Cu-catalyzed alkyne-azide cycloaddition. Metallation with Ni(II).
Covalent tethering of P(Ph)2N(C6H4C≡CH)2 ligands (P(Ph)2N(C6H4C≡CH)2 = 1,5-di-(4-ethynylphenyl)-3,7-diphenyl-1,5-diaza-3,7-diphosphacyclooctane) to planar, azide-terminated glassy carbon electrode surfaces has been accomplished using a Cu(I)-catalyzed alkyne-azide cycloaddition (CuAAC) coupling reaction, using a BH3←P protection-deprotection strategy. Deprotected, surface-confined ligands were ...
متن کاملCarbohydrate and protein immobilization onto solid surfaces by sequential Diels-Alder and azide-alkyne cycloadditions.
We demonstrate the applicability of sequential Diels-Alder and azide-alkyne [3 + 2] cycloaddition reactions (click chemistry) for the immobilization of carbohydrates and proteins onto a solid surface. An alpha,omega-poly(ethylene glycol) (PEG) linker carrying alkyne and cyclodiene terminal groups was synthesized and immobilized onto an N-(epsilon-maleimidocaproyl) (EMC)-functionalized glass sli...
متن کاملClick chemistry in mesoporous materials: functionalization of porous silicon rugate filters.
In this paper we report the use of the optical properties of porous silicon photonic crystals, combined with the chemical versatility of acetylene-terminated SAMs, to demonstrate the applicability of "click" chemistry to mesoporous materials. Cu(I)-catalyzed alkyne-azide cycloaddition reactions were employed to modify the internal pore surfaces through a two-step hydrosilylation/cycloaddition p...
متن کاملApplication of Cu(I)-catalyzed azide–alkyne cycloaddition for the design and synthesis of sequence specific probes targeting double-stranded DNA
Efficient protocols based on Cu(I)-catalyzed azide-alkyne cycloaddition were developed for the synthesis of conjugates of pyrrole-imidazole polyamide minor groove binders (MGB) with fluorophores and with triplex-forming oligonucleotides (TFOs). Diverse bifunctional linkers were synthesized and used for the insertion of terminal azides or alkynes into TFOs and MGBs. The formation of stable tripl...
متن کامل