Glomalin is a soil protein resembling heat shock protein (HSP) 60 and exerting high affinity to metals, causing retention of water in the environment and improving mechanical stability of soil. Currently, glomalin is determined in the soil or other samples by combination of autoclaving extraction and total protein determination typically by the Bradford method. In this paper, a piezoelectric biosensor was prepared to determine glomalin in a label-free measurement. The biosensor contained antibodies immobilized on quartz crystal microbalance (QCM), and the recognition layer was stabilized by iron oxide nanoparticles. The assay was tested on real soil samples and compared with the standard Bradford assay. Limit of detection of the assay was equal to 2.4 µg/g for a soil extract with a volume of 50 µl. The assay takes approximately half of an hour and was fully correlated to the Bradford assay. The biosensor had significant advantages than the other methods: it worked in a label-free mode and was fully applicable for practical samples.

Faculty of AgriSciences Mendel University in Brno Zemedelska 1 613 00 Brno Czech Republic

Faculty of Military Health Sciences University of Defence Trebesska 1575 CZ 500 01 Hradec Kralove Czech Republic

Zobrazit více v PubMed

Wang Q., Lu H., Chen J., et al. Interactions of soil metals with glomalin-related soil protein as soil pollution bioindicators in mangrove wetland ecosystems. Science of the Total Environment. 2020;709:p. 136051. doi: 10.1016/j.scitotenv.2019.136051. PubMed DOI

Vlcek V., Pohanka M. Glomalin-an interesting protein part of the soil organic matter. Soil and Water Research. 2020;15:67–74.

Wang F., Adams C. A., Yang W., Sun Y., Shi Z. Benefits of arbuscular mycorrhizal fungi in reducing organic contaminant residues in crops: implications for cleaner agricultural production. Critical Reviews in Environmental Science and Technology. 2020;50(15):p. 1580. doi: 10.1080/10643389.2019.1665945. DOI

Janos D. P., Garamszegi S., Beltran B. Glomalin extraction and measurement. Soil Biology and Biochemistry. 2008;40(3):728–739. doi: 10.1016/j.soilbio.2007.10.007. DOI

Moragues-Saitua L., Merino-Martín L., Stokes A., Staunton S. Towards meaningful quantification of glomalin-related soil protein (GRSP) taking account of interference in the coomassie blue (Bradford) assay. European Journal of Soil Science. 2019;70(4):727–735. doi: 10.1111/ejss.12698. DOI

Jorge-Araújo P., Quiquampoix H., Matumoto-Pintro P. T., Staunton S. Glomalin-related soil protein in French temperate forest soils: interference in the bradford assay caused by co-extracted humic substances. European Journal of Soil Science. 2015;66(2):311–319. doi: 10.1111/ejss.12218. DOI

Reyna D. L., Wall L. G. Revision of two colorimetric methods to quantify glomalin-related compounds in soils subjected to different managements. Biology and Fertility of Soils. 2014;50(2):395–400. doi: 10.1007/s00374-013-0834-2. DOI

Koide R. T., Peoples M. S. Behavior of bradford-reactive substances is consistent with predictions for glomalin. Applied Soil Ecology. 2013;63:8–14. doi: 10.1016/j.apsoil.2012.09.015. DOI

Wright S. F., Jawson L. A pressure cooker method to extract glomalin from soils. Soil Science Society of America Journal. 2001;65(6):1734–1735. doi: 10.2136/sssaj2001.1734. DOI

Malekzadeh E., Aliasgharzad N., Majidi J., Aghebati-Maleki L., Abdolalizadeh J. Cd-induced production of glomalin by arbuscular mycorrhizal fungus (rhizophagus irregularis) as estimated by monoclonal antibody assay. Environmental Science and Pollution Research. 2016;23(20):20711–20718. doi: 10.1007/s11356-016-7283-z. PubMed DOI

Liu Y., Pan M., Wang W., et al. Plasmonic and photothermal immunoassay via enzyme-triggered crystal growth on gold nanostars. Analytical Chemistry. 2019;91(3):2086–2092. doi: 10.1021/acs.analchem.8b04517. PubMed DOI

Yu Z., Tang Y., Cai G., Ren R., Tang D. Paper electrode-based flexible pressure sensor for point-of-care immunoassay with digital multimeter. Analytical Chemistry. 2019;91(2):1222–1226. doi: 10.1021/acs.analchem.8b04635. PubMed DOI

Luo Z., Qi Q., Zhang L., Zeng R., Su L., Tang D. Branched polyethylenimine-modified upconversion nanohybrid-mediated photoelectrochemical immunoassay with synergistic effect of dual-purpose copper ions. Analytical Chemistry. 2019;91(6):4149–4156. doi: 10.1021/acs.analchem.8b05959. PubMed DOI

Ma H., Zhao Y., Liu Y., et al. A compatible sensitivity enhancement strategy for electrochemiluminescence immunosensors based on the biomimetic melanin-like deposition. Analytical Chemistry. 2017;89(24):13049–13053. doi: 10.1021/acs.analchem.7b04397. PubMed DOI

Ren R., Cai G., Yu Z., Zeng Y., Tang D. Metal-polydopamine framework: an innovative signal-generation tag for colorimetric immunoassay. Analytical Chemistry. 2018;90(18):11099–11105. doi: 10.1021/acs.analchem.8b03538. PubMed DOI

Chen Z., Liu Y., Wang Y., Zhao X., Li J. Dynamic evaluation of cell surface N-glycan expression via an electrogenerated chemiluminescence biosensor based on concanavalin a-integrating gold-nanoparticle-modified Ru (bpy)32+-doped silica nanoprobe. Analytical Chemistry. 2013;85(9):4431–4438. doi: 10.1021/ac303572g. PubMed DOI

Pohanka M. The piezoelectric biosensors: principles and applications, a review. International Journal of Electrochemical Science. 2017;12:496–506. doi: 10.20964/2017.01.44. DOI

Pohanka M. Overview of piezoelectric biosensors, immunosensors and DNA sensors and their applications. Materials. 2018;11(3):p. 448. doi: 10.3390/ma11030448. PubMed DOI PMC

Pohanka M. Piezoelectric biosensor for the determination of tumor necrosis factor alpha. Talanta. 2018;178:970–973. doi: 10.1016/j.talanta.2017.10.031. PubMed DOI

Pirich C. L., De Freitas R. A., Torresi R. M., Picheth G. F., Sierakowski M. R. Piezoelectric immunochip coated with thin films of bacterial cellulose nanocrystals for dengue detection. Biosensors and Bioelectronics. 2017;92:47–53. doi: 10.1016/j.bios.2017.01.068. PubMed DOI

Wang Z. L. Progress in piezotronics and piezo-phototronics. Advanced Materials. 2012;24(34):4632–4646. doi: 10.1002/adma.201104365. PubMed DOI

Wright S. F., Upadhyaya A. Extraction of an abundant and unusual protein from soil and comparison with hyphal protein of arbuscular mycorrhizal fungi. Soil Science. 1996;161(9):575–586. doi: 10.1097/00010694-199609000-00003. DOI

Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry. 1976;72(1-2):248–254. doi: 10.1016/0003-2697(76)90527-3. PubMed DOI

Nautiyal P., Rajput R., Pandey D., Arunachalam K., Arunachalam A. Role of glomalin in soil carbon storage and its variation across land uses in temperate himalayan regime. Biocatalysis and Agricultural Biotechnology. 2019;21:p. 101311. doi: 10.1016/j.bcab.2019.101311. DOI

Huang X., Chen Q., Pan W., Hu J., Yao Y. Assessing the mass sensitivity for different electrode materials commonly used in quartz crystal microbalances (QCMs) Sensors. 2019;19(18):p. 3968. doi: 10.3390/s19183968. PubMed DOI PMC

Kim J., Urchaga P., Baranton S., Coutanceau C., Jerkiewicz G. Interfacial structure of atomically flat polycrystalline Pt electrodes and modified sauerbrey equation. Physical Chemistry Chemical Physics. 2017;19(33):21955–21963. doi: 10.1039/c7cp02528a. PubMed DOI

Heller G. T., Mercer-Smith A. R., Johal M. S. Quartz microbalance technology for probing biomolecular interactions. Methods in Molecular Biology. 2015;1278:153–164. doi: 10.1007/978-1-4939-2425-7_9. PubMed DOI

Kang Q., Shen Q., Zhang P., Wang H., Sun Y., Shen D. Unfound associated resonant model and its impact on response of a quartz crystal microbalance in the liquid phase. Analytical Chemistry. 2018;90(4):2796–2804. doi: 10.1021/acs.analchem.7b04906. PubMed DOI

Huang X., Bai Q., Hu J., Hou D. A practical model of quartz crystal microbalance in actual applications. Sensors (Basel) 2017;17:p. 1785. doi: 10.3390/s17071476. PubMed DOI PMC

Anderson N. L., Anderson N. G. The human plasma proteome. Molecular & Cellular Proteomics. 2002;1(11):845–867. doi: 10.1074/mcp.r200007-mcp200. PubMed DOI

Bal W., Sokolowska M., Kurowska E., Faller P. Binding of transition metal ions to albumin: sites, affinities and rates. Biochimica et Biophysica Acta (BBA) 2013;1830(12):5444–5455. doi: 10.1016/j.bbagen.2013.06.018. PubMed DOI

Zurawska-Plaksej E., Rorbach-Dolata A., Wiglusz K., Piwowar A. The effect of glycation on bovine serum albumin conformation and ligand binding properties with regard to gliclazide. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy. 2018;189:625–633. doi: 10.1016/j.saa.2017.08.071. PubMed DOI

Nishi K., Yamasaki K., Otagiri M. Serum albumin, lipid and drug binding. Subcellular Biochemistry. 2020;94:383–397. doi: 10.1007/978-3-030-41769-7_15. PubMed DOI

Duce C., Bramanti E., Ghezzi L., et al. Interactions between inorganic pigments and proteinaceous binders in reference paint reconstructions. Dalton Transactions. 2013;42(17):5975–5984. doi: 10.1039/c2dt32203j. PubMed DOI

Villa P., Pollarolo L., Degano I., et al. A milk and ochre paint mixture used 49,000 years ago at sibudu, South Africa. PLoS One. 2015;10 doi: 10.1371/journal.pone.0131273.e0131273 PubMed DOI PMC

Wadley L. Putting ochre to the test: replication studies of adhesives that may have been used for hafting tools in the middle stone age. Journal of Human Evolution. 2005;49(5):587–601. doi: 10.1016/j.jhevol.2005.06.007. PubMed DOI

Pohanka M., Vlcek V. Assay of glomalin using a quartz crystal microbalance biosensor. Electroanalysis. 2018;30(3):453–458. doi: 10.1002/elan.201700772. DOI