Title :
Investigation of the binding affinity of C-terminal domain of SARS coronavirus nucleocapsid protein to nucleotide using AlGaN/GaN high electron mobility transistors
Author :
You-Ren Hsu ; Geng-Yen Lee ; Jen-Inn Chyi ; Chung-ke Chang ; Chih-Cheng Huang ; Chen-Pin Hsu ; Tai-Huang Huang ; Fan Ren ; Yu-Lin Wang
Author_Institution :
Inst. of Nanoengineering & Microsyst., Nat. Tsing Hua Univ., Hsinchu, Taiwan
Abstract :
In this article, we used AlGaN/GaN high electron mobility transistors (HEMTs) to construct the first label-free semiconductor-sensor-based binding assay to our knowledge. Our results suggested that the nucleotide-c terminal domain of SARS coronavirus (SARS-CoV) nucleocapsid protein interaction is a two-step binding event with two dissociation constants (Kd1 = 0.052 nM, and Kd2 = 51.24nM) extracted by using the modified two-binding-site Langmuir isotherm equation proposed here. We found that there were at least two protein binding sites on the specific 41-base SARS-CoV double-stranded DNA (dsDNA) genome (29,580-29621) conjugated with a 20-mer poly-dT tail. This result presented a high binding affinity is comparable with the antibody-antigen reaction, and suggested this designed dsDNA could be treated as an aptamer for SARS-CoV N protein capture.
Keywords :
III-V semiconductors; aluminium compounds; biological techniques; biomedical electronics; gallium compounds; high electron mobility transistors; proteomics; AlGaN-GaN; C-terminal domain; HEMT; Langmuir isotherm equation; SARS coronavirus nucleocapsid protein; SARS-CoV N protein capture; SARS-CoV nucleocapsid protein interaction; antibody-antigen reaction; binding affinity; binding assay; double stranded DNA genome 29580-29621; high electron mobility transistors; label-free semiconductor sensor; nucleotide-c terminal domain; Aluminum gallium nitride; Equations; Gallium nitride; HEMTs; MODFETs; Mathematical model; Proteins;
Conference_Titel :
Sensors, 2012 IEEE
Conference_Location :
Taipei
Print_ISBN :
978-1-4577-1766-6
Electronic_ISBN :
1930-0395
DOI :
10.1109/ICSENS.2012.6411277