title: A one-pot strategy for the detection of proteins based on sterically and allosterically tunable hybridization chain reaction.
abstract: In this work, we report a facile one-pot strategy for protein detection based on sterically and allosterically tunable hybridization chain reaction (HCR). In our strategy, DNA hairpins H1 and H2 are dual-labeled with pyrene moieties through a six-carbon-atom spacer at each end; and a single-stranded DNA primer is designed to contain two small molecules near each end. In the absence of target protein, the primer can trigger HCR events between alternating H1 and H2 hairpins to form a nicked double-helix. As a result, the pyrene excimers are formed to emit at approximately 485nm. On the contrary, upon binding of the specific target protein onto the primer through the protein-small molecule interaction, the HCR will be inhibited due to the steric and allosteric effect. The changes of the fluorescent signals of pyrene excimers are in response to the concentration of target protein, so that the detection of protein can be realized. We have demonstrated the feasibility of this strategy by using streptavidin (SA) and folate receptor (FR) as model targets. Results show that both of them can be well detected with a detection limit of 1.07nM and 2.7nM, respectively. The developed method for protein assay is flexible, so we infer that the one-pot strategy holds great potential for the detection of other proteins.
authors.affiliation: Laboratory of Biosensing Technology, School of Life Sciences, Shanghai University, Shanghai 200444, China.
authors.forename: Fangfang
authors.initials: F
authors.lastname: Ban
authors.affiliation: Laboratory of Biosensing Technology, School of Life Sciences, Shanghai University, Shanghai 200444, China; State Key Laboratory of Pharmaceutical Biotechnology, Department of Biochemistry, Nanjing University, Nanjing 210093, China.
authors.forename: Hai
authors.initials: H
authors.lastname: Shi
authors.affiliation: State Key Laboratory of Pharmaceutical Biotechnology, Department of Biochemistry, Nanjing University, Nanjing 210093, China.
authors.forename: Chang
authors.initials: C
authors.lastname: Feng
authors.affiliation: Laboratory of Biosensing Technology, School of Life Sciences, Shanghai University, Shanghai 200444, China.
authors.forename: Xiaoxia
authors.initials: X
authors.lastname: Mao
authors.affiliation: Department of Oncology, the First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China. Electronic address: ym.yin@hotmail.com.
authors.forename: Yongmei
authors.initials: Y
authors.lastname: Yin
authors.affiliation: Laboratory of Biosensing Technology, School of Life Sciences, Shanghai University, Shanghai 200444, China. Electronic address: xiaolizhu@shu.edu.cn.
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Zhu
mesh_terms: D015342:DNA Probes / Q000737:chemistry*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D017404:In Situ Hybridization, Fluorescence / Q000295:instrumentation*; D040081:Protein Array Analysis / Q000295:instrumentation*; D011506:Proteins / Q000032:analysis* / Q000235:genetics; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D015342:DNA Probes; D011506:Proteins
keywords: Allosteric effect; Hybridization chain reaction; Protein; Small molecule; Steric effect