title: A pH Indicator-linked Immunosorbent assay following direct amplification strategy for colorimetric detection of protein biomarkers.
abstract: A new pH indicator-linked immunosorbent assay (PILISA) reached pg/mL sensitivity based on pH indicator molecules loaded carbon nitride nanosheets as signal enhancer has been developed for colorimetric detection of protein biomarkers. As the secondary antibody binds to the carbon nitride nanosheets, the carbon nitride nanosheets and pH indicator complex as the signal amplification platform for colour change by detecting absorbance of pH indicator. The colour change was resulted from the releasing of pH indicator molecules from carbon nitride nanosheets triggered by alkali solution (AS). In this novel PILISA, the intensity absorbance of pH indicator is proportional to the concentration of the disease marker. The outstanding detection performance of the PILISA can be attributed to the following reasons: (1) ultrathin carbon nitride nanosheets with a larger surface area could adsorb abundant phenolphthalein (PP) molecules through hydrophobic interactions as well as the resulted PP anions can be free easily released into aqueous solution, leading to an obvious allochroic response; (2) the signal intensity is precisely determined by the amount of PP molecules loading onto the carbon nitride nanosheets surface, which ensures simple, low-cost and stable colorimetric detection. As expected, this new PILISA method offered an enzyme-free approach followed enzyme-linked immunosorbent assay format, which showed great promising potential as an innovative robust assay method for practical clinical applications.
authors.affiliation: School of Chemistry and Chemical Engineering, and School of Biological Science and Technology, University of Jinan, Jinan 250022, China.
authors.forename: Fengying
authors.initials: F
authors.lastname: Shao
authors.affiliation: School of Chemistry and Chemical Engineering, and School of Biological Science and Technology, University of Jinan, Jinan 250022, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Jiao
authors.affiliation: School of Chemistry and Chemical Engineering, and School of Biological Science and Technology, University of Jinan, Jinan 250022, China.
authors.forename: Luyang
authors.initials: L
authors.lastname: Miao
authors.affiliation: School of Chemistry and Chemical Engineering, and School of Biological Science and Technology, University of Jinan, Jinan 250022, China.
authors.forename: Qin
authors.initials: Q
authors.lastname: Wei
authors.affiliation: School of Chemistry and Chemical Engineering, and School of Biological Science and Technology, University of Jinan, Jinan 250022, China. Electronic address: lihecd@gmail.com.
authors.forename: He
authors.initials: H
authors.lastname: Li
mesh_terms: D015415:Biomarkers / Q000737:chemistry*; D015374:Biosensing Techniques / Q000379:methods*; D003124:Colorimetry; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007164:Immunosorbents / Q000737:chemistry*; D053758:Nanoparticles / Q000737:chemistry; D009570:Nitriles / Q000737:chemistry; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification*
chemical_list: D015415:Biomarkers; D007164:Immunosorbents; D009570:Nitriles; D011506:Proteins; C011206:cyanogen
keywords: Carbon nitride nanosheets; Colorimetric detection; Protein biomarkers; pH indicator; pH indicator-linked immunosorbent assay