Integrated Tyramide and Polymerization-Assisted Signal Amplification for a Highly-Sensitive Immunoassay

• Published in: Analytical chemistry (Washington) Vol. 84; no. 24; pp. 10737 - 10744
• Main Authors: Yuan, Liang, Xu, Lingling, Liu, Songqin
• Format: Journal Article
• Language: English
• Published: Washington, DC American Chemical Society 18-12-2012
• Subjects: Analytical chemistry; Chemistry; Electrochemical methods; Electrodes; Exact sciences and technology; Humans; Immunoassay; Immunoassay - methods; Immunoglobulin G - chemistry; Immunoglobulins; Miscellaneous; Polymerization; Proteins; Quantum dots; Tyramine - chemistry; Fold; Hydrogen peroxide; IgG; Immobilization; Voltammetry; Immunological method; Target; Signal; Loading; Detection limit; Peroxidase; Coupling; Human; Immunoglobulins; Enzyme; Long chain; Ultratrace compound; Polymerization; Electrochemiluminescence; Protein; Antigen; Gene amplification; Sensitivity; Peroxidases; Transfer; Strategy; Feasibility; Oxidoreductases; Detection; Application; Comparative study; Square wave
• ISSN: 0003-2700; 1520-6882
• DOI: 10.1021/ac302439v

A novel strategy for ultrasensitive detection of model protein based on the integration of tyramide signal amplification (TSA) and polymerization-assisted signal amplification was proposed. The surface-initiated atom transfer radical polymerization (SI-ATRP) of glycidyl methacrylate (GMA) was triggered by the initiator-coupled protein immobilized on the electrode surface through sandwiched immunoreactions. Growth of long chain polymeric materials provided numerous epoxy groups for subsequent coupling of horseradish peroxidase (HRP), which in turn significantly increased the loading of quantum dots (QDs) labeled tyramide in the presence of hydrogen peroxide. As a result, electrochemiluminescence (ECL) and square-wave voltammetric (SWV) measurements showed 9.4- and 10.5-fold increase in detection signal in comparison with the unamplified method, respectively. To demonstrate the feasibility of this approach, human immunoglobulin G antigen (IgG) as a model target protein was employed and the detection limits were 0.73 and 0.09 pg mL–1 for ECL and SWV, respectively. The results showed that sensitivity of the presented immunoassay significantly increased by one-order of magnitude and offered great application promises in providing a sensitive, specific, and potent method for biological detection.