A fluorometric and colorimetric method for determination of trypsin by exploiting the gold nanocluster-induced aggregation of hemoglobin-coated gold nanoparticles
- 66 Downloads
A dual-signal assay is described for the determination of trypsin based on the use of gold nanoparticles (AuNPs) that aggregate in the presence of gold nanoclusters (AuNCs) due to electrostatic interaction. This is accompanied by a color change from red to blue. However, if hemoglobin (Hb) is present in the solution, it will attach to the surface of AuNPs, thus preventing aggregation. The Hb-coated AuNPs quench the fluorescence of AuNCs. Trypsin can hydrolyze Hb and destroy the protective coating of Hb on the AuNPs. As a result, AuNP aggregation will occur after the addition of AuNCs, and the blue fluorescence of the AuNCs with 365 nm excitation and 455 nm maximum emission peak is recovered. Thus, trypsin can be determined by measurement of fluorescence emission intensity. Additionally, trypsin can be determined by the maximum absorption peak wavelength between 530 nm and 610 nm. Fluorescence increases linearly in the 10–2500 ng⋅mL−1 concentration range, and absorbance in the 20–2000 ng·mL−1 concentration range. The limits of detection are 4.6 ng·mL−1 (fluorometry) and 8.4 ng·mL−1 (colorimetry), respectively. The assay is sensitive and selective, and can be applied to the determination of trypsin in serum.
KeywordsDual-signal assay Fluorescence resonance energy transfer Hemoglobin coated gold nanoparticles Fluorescence Colorimetry Electrostatic interaction Serum analysis
This work was supported by a grant from the Two-Way Support Programs of Sichuan Agricultural University (Project No.03570113), the Education Department of Sichuan Provincial, P. R. China (Grant No. 16ZA0039), National Natural Science Foundation of China (Grant No. 11404358).
Compliance with ethical standards
The author(s) declare that they have no competing interests.
- 16.Cheng C, Chen H-Y, Wu C-S, Meena JS, Simon T, Ko F-H (2016) A highly sensitive and selective cyanide detection using a gold nanoparticle-based dual fluorescence–colorimetric sensor with a wide concentration range. Sensor Actuat B-Chem 227:283–290. https://doi.org/10.1016/j.snb.2015.12.057 CrossRefGoogle Scholar
- 17.Liu T, Li N, Dong JX, Zhang Y, Fan YZ, Lin SM, Luo HQ, Li NB (2017) A colorimetric and fluorometric dual-signal sensor for arginine detection by inhibiting the growth of gold nanoparticles/carbon quantum dots composite. Biosens Bioelectron 87:772–778. https://doi.org/10.1016/j.bios.2016.08.098 CrossRefPubMedGoogle Scholar
- 18.Ma X, Gao L, Tang Y, Miao P (2017) Gold nanoparticles-based DNA logic gate for miRNA inputs analysis coupling Strand displacement reaction and hybridization chain reaction. Part Part Syst Charact:1700326Google Scholar
- 22.Garabagiu S (2011) A spectroscopic study on the interaction between gold nanoparticles and hemoglobin. Mater Res Bull 46(12):2474–2477. https://doi.org/10.1016/j.materresbull.2011.08.032 CrossRefGoogle Scholar
- 25.She W, Luo K, Zhang C, Wang G, Geng Y, Li L, He B, Gu Z (2013) The potential of self-assembled, pH-responsive nanoparticles of mPEGylated peptide dendron–doxorubicin conjugates for cancer therapy. Biomaterials 34(5):1613–1623. https://doi.org/10.1016/j.biomaterials.2012.11.007 CrossRefPubMedGoogle Scholar
- 32.J M A (1981) Serum trypsin levels in acute pancreatic and non-pancreatic abdominal conditions. Postgrad Med J 57(666)Google Scholar
- 34.Ariga GG, Naik PN, Chimatadar SA, Nandibewoor ST (2017) Interactions between epinastine and human serum albumin: investigation by fluorescence, UV–vis, FT–IR, CD, lifetime measurement and molecular docking. J Mol Struct 1137:485–494. https://doi.org/10.1016/j.molstruc.2016.12.066 CrossRefGoogle Scholar