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Development of a new and simple method for the detection of histidine-tagged proteins based on thionine-chitosan/gold nanoparticles/horseradish peroxidase

Authors
  • Ren, Ruijuan1, 2
  • Lu, Dingqiang1, 2
  • Pang, Guangchang1, 2
  • 1 Tianjin University of Commerce, Tianjin, 300314, China , Tianjin (China)
  • 2 Tianjin key laboratory of food biotechnology, Tianjin, 300314, China , Tianjin (China)
Type
Published Article
Journal
Biomedical Microdevices
Publisher
Springer-Verlag
Publication Date
Jan 02, 2020
Volume
22
Issue
1
Identifiers
DOI: 10.1007/s10544-019-0464-z
Source
Springer Nature
Keywords
License
Yellow

Abstract

In the current study, an electrochemical biosensing signal amplification system was utilized with thionine-chitosan-gold nanoparticles (Chit-GNPs) that absorbed horseradish peroxidase (HRP) and anti-His tagged protein monoclonal antibody derived from Balb/c mice. In addition, transmission electron microscopy (TEM) was used to characterize the nanogold solution and atomic force microscopy (AFM) was used to characterize the sensor assembly. To evaluate the quality of the immunosensor, the amperometric I-t curve method was applied to determine His-IL23 in PBS. The results indicated that the response current exhibited an optimal linear correlation with the His-IL23 concentration that ranged from 0.01 to 103 ng/ml. The lowest detection limit was noted at 3.3 pg/ml (S/N = 3). The linear equation was deduced as follows: △I = 0.02lgC + 0.037 (R2 = 0.9628). Moreover, it was validated with high sensitivity, reproducibility and rapid response. Apparently, the immunosensor may be a very useful tool for the detection and quantification of His-tagged proteins. In addition, the signal amplification system can be used for the preparation of other immunosensors and to assist in bioassays.

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