Nashrieh Shimi va Mohandesi Shimi Iran

Nashrieh Shimi va Mohandesi Shimi Iran

Functionalization of Graphene Oxide with the Heavy Chain of Antibody and Its Interaction with the Labeled Light Chain for the Detection of Bone Gala Protein Based on Fluorescence Spectroscopy

Document Type : Research Article

Authors
Javad Gholami 1
Mehrdad Manteghian 2
1 Department of Applied Chemistry, Faculty of Basic Sciences, Malayer University, Malayer, I.R.IRAN
2 Faculty of Chemical Engineering, Tarbiat Modares University, Tehran, I.R.IRAN
Abstract
Biomarkers are biological structures, such as proteins, whose measurement is highly important for the early diagnosis of serious diseases like cancer. Graphene oxide due to its unique properties has recently attracted considerable attention for the detection of specific biological samples based on luminescence spectroscopy. In this study, graphene oxide was synthesized and analyzed using X-ray photoelectron spectroscopy (XPS). Subsequently, graphene oxide in the solution phase was functionalized with the heavy chain of a bone gla protein antibody in the presence of chemical reagents and biologically identified. The interaction between the functionalized graphene oxide with the heavy chain of the antibody and the labeled light chain of the antibody led to a quenching of fluorescence emission. Upon adding bone gala protein to the solution containing the functionalized graphene oxide and the labeled light chain, based on the immunosandwich mechanism, the fluorescence emission intensity was recovered and its used for the detection of bone gla protein. This signal allowed the detection of the biomarker in the nanomolar concentration range.
Keywords
Graphene oxide
Fluorescence
Immunosandwich
Bone gla protein
Antibody chain

Subjects

[1] Hee J.J., Quenchbodies That Enable One-Pot Detection of Antigens. A Structural Perspective. Bioengineering, 10: 1262 (2023).
[2] Fritzenschaft L., Boehm F., Rothenbacher D., Denkinger M., Dallmeier D., Association of Blood Biomarkers with Frailty–A Mapping Review. Ageing Research Reviews, 109: 102761 (2025).
[3] Sarhadi V.K., Armengol G., Molecular Biomarkers in Cancer. Biomolecules, 12: 1021(2022).
[4] Jong P.K., Seon H.K., Ueda H., Hee J.J., Generation of Q-Bead Against Bone Gla Protein with Simplified Preparation Steps. Journal of Immunological Methods, 516: 113471 (2023).
[5] Price P., Parthemore J., Deftos L., New Biochemical Marker for Bone metabolismNew Biochemical Marker for Bone Metabolism. Measurement by Radioimmunoassay of Bone GLA Protein in the Plasma of Normal Subjects and Patients with Bone Disease. Journal of Clinical Investigation, 66: 878-887 (1980).
[6] Idelevich A., Rais Y., Monsonego-Ornan E., Bone Gla Protein Increases HIF-1α–Dependent Glucose Metabolism and Induces Cartilage and Vascular Calcification. Arteriosclerosis, thrombosis, and vascular biology, 31: 55-62 (2011).
[7] Azria M., The Value of Biomarkers in Detecting Alterations in Bone Metabolism. Calcified tissue international, 45: 7-13 (1989).
[8] Deeksha N., Utkarsh J., Emerging Trends in Fluorescent and Quenching Nanomaterials for Viral Detection. Innovations in Biological and Chemical sensing,Talanta, 11: 100430 (2025).
[9] Cho Y., Choi Y., Jang Y., Seong H., Nanomaterial-Enhanced Biosensing: Mechanisms and Emerging Applications. Adv. Healthcare Mater, 2500189 (2025).
[10] He X., Zhang L., Qi H., Yu P., Fei J., Mao L., Improving the Fluorescence Detection Limit with Positively Charged Carbon Nanostructures as a Low Background Signal Platform. Analyst, 139: 2114-2120 (2014).
[11] Norouzi F., Pourmadadi M., Yazdian F., Khoshmaram K., Mohammadnejad J., Sanati MH., Chogan F., Rahdar A., Baino F., PVA-Based Nanofibers Containing Chitosan Modified with Graphene Oxide and Carbon Quantum Dot-Doped TiO2 Enhance Wound Healing in a Rat Model. Journal of Functional Biomaterials, 13: 300(2022).
[12] Dong H., Gao W., Yan F., Ji H., . Ju H., Fluorescence Resonance Energy Transfer Between Quantum Dots and Graphene Oxide for Sensing Biomolecules. Analytical chemistry, 82: 5511-5517 (2010).
[13] Jang H., Kim Y.K., Kwon H.M., Yeo W.S., Kim D.E., Min D.H., A Graphene‐Based Platform for the Assay of Duplex‐DNA Unwinding by Helicase. Angewandte Chemie, 122: 5703-5707 (2010).
[14] Song Y., Yang X., Li Z., Zhao Y., Fan A., Label-Free Chemiluminescent ATP Aptasensor Based on Graphene Oxide and an Instantaneous Derivatization of Guanine Bases. Biosensors and Bioelectronics, 51: 232-237 (2014).
[15] Zamani M., Pourmadadi M., Ebrahimi S. S., Yazdian F., Shayeh J. S., A Novel Labeled and Label-Free Dual Electrochemical Detection of Endotoxin Based on Aptamer-Conjugated Magnetic Reduced Graphene Oxide-Gold Nanocomposite. Journal of Electroanalytical Chemistry, 908: 116116 (2022).
[16] Jung J.H., Cheon D.S., Liu F., Lee K.B., Seo T.S., A Graphene Oxide Based Immuno‐Biosensor for Pathogen Detection. Angewandte Chemie International Edition, 49: 5708-5711 (2010).
[17] Chen J.L., Yan X.-P., Meng K., Wang S.F., Graphene Oxide Based Photoinduced Charge Transfer Label-Free Near-Infrared Fluorescent Biosensor for Dopamine. Analytical chemistry, 83: 8787–8793 (2011).
[18] Astrid S., Helmut S., Roland K.O.S., Interrelations Between Essential Metal Ions and Human Diseases. springer, (2013).
[19] Gholami J., Manteghian M., Badiei A., Javanbakht M., Ueda H., Label Free Detection of Vitamin B12 Based on Fluorescence Quenching of Graphene Oxide Nanolayer. Fuller. Nanotub. Carbon Nanostructure,  23: 878-884 (2015).
[20] Gholami J., Manteghian M., Badiei A., Ueda H., Javanbakht M., N-Butylamine Functionalized Graphene Oxide for Detection of Iron(III) by Photoluminescence Quenching. Luminescence, 31: 229–235 (2016).
[21] Hummers W. S. J., Offeman R.E., Preparation of Graphitic Oxide. Journal of the American Chemical Society, 80: 1339 (1958).
[22] Roy S., Soin N., Bajpai R., Misra D.S., McLaughlinb J.A., Roy S.S., Graphene Oxide for Electrochemical Sensing Applications. Journal of Materials Chemistry, 21: 14725-14731 (2011).
[23] Dreyer D.R., Park S., Bielawski C.W., Ruoff R.S., The Chemistry of Graphene Oxide, Chemical Society Reviews, 39: 228-240 (2010).
[24] Tao C.a., Wang J., Qin S., Lv Y., Long Y., Zhu H., Jiang Z., Fabrication of pH-Sensitive Graphene Oxide–Drug Supramolecular Hydrogels as Controlled Release Systems. Journal of Materials Chemistry, 22: 24856-24861 (2012).
[25] Hasan S., Dong J., Hara Y., Morizane Y., Shibasaki F., Ueda H., Protein-Based Open Sandwich Immuno-PCR for Sensitive Detection of Small Biomarkers. Analytical Sciences, 29: 871-876 (2013).