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Updated: Jul 3, 2026

Aptamer-Based Target Detection Facilitated by a 3-Stage G-Quadruplex Isothermal Exponential Amplification Reaction
Published on: October 6, 2022
Conjugation of DNA with protein using His-tag chemistry and its application to the aptamer-based detection system
Josui Shimada1, Tatsuo Maruyama, Takuya Hosogi
1Department of Applied Chemistry, Graduate School of Engineering, Center for Future Chemistry, Kyushu University, 744 Moto-oka, Fukuoka, 819-0395, Japan.
Abstract:
We propose a novel method to prepare a DNA-protein conjugate using histidine-tag (His-tag) chemistry. Oligo-DNA was modified with nitrilotriacetate (NTA), which has high affinity to a His-tag on recombinant protein via the complexation of Ni(2+). Investigations using a microplate which displayed a complementary DNA-strand revealed that a NTA-modified DNA-protein conjugate was formed and immobilized in the presence of Ni(2+) on the microplate. We then adopted alkaline phosphatase (AP) as a model protein, and application of the DNA-AP conjugate was demonstrated in a thrombin aptamer-based detection system with a detection limit of approximately 10 nM.
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