Poly Oligonucleotides Antifouling Interface with Highly Specific Response of Multi-Biomarkers in Serum: Anti-Fouling Mechanisms and Influencing Factors

Abstract

Quantifying trace markers in clinical samples with electrochemical impedance spectroscopy (EIS) biosensors is limited mainly by ubiquitous biofouling because traditional antifouling coatings suffer from low stability and unsatisfactory antifouling capability etc. Herein, a simple and effective strategy was constructed to achieve a highly specific EIS biosensing platform for quantifying protein and DNA in serum by using single-stranded oligonucleotides (ssDNA) as antifouling coating. To achieve the optimized antifouling capability of ssDNA coating, the influence factors of ssDNA structure are investigated by combining electrochemical experiments with theoretical calculations. The results demonstrated that the chemical adsorption of adenine on the Au surface is stronger than that of the other three bases, which endows the poly adenine-ssDNA (polyAn) coating with outstanding antifouling capability. Improtantly, we found that the capture ligand structure and ssDNA length remarkably affected the antifouling capability of polyAn because of steric hindrance. As a proof of concept, EIS biosensors based on polyAn antifouling coating were developed to detect target DNA and nuclear factor kappa b p65 (NF-kB p65), respectively, exhibiting outstanding capability of detecting DNA and NF-kB p65 in human serum with low interference and good accuracy. Overall, the presented work may contribute to a deeper understanding of the influence factors of ssDNA-based antifouling coating and holds great insight into the construction of highly specific biosensors for quantifying disease markers in clinical samples.

Note:
Funding Information: The authors are grateful to the National Natural Science Foundation of China (21804014, 41272321), the Science and Technology Research Program of Chongqing Municipal Education Commission (KJQN201800827, KJQN201900837), Key Platform Open Fund of Chongqing Technology and Business University (KFJJ2019083).

Conflict of Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Keywords: Electrochemical impedance biosensors, antifouling, Poly oligonucleotides, Poly adenine-ssDNA, Nuclear factor kappa b p65, Clinical samples