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Functionalized Graphene Quantum Dots (GQDs) based Label-Free Optical Fluorescence Sensor for CD59 Antigen Detection and Cellular Bioimaging

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Abstract

Cluster of differentiation (CD59), a cell surface glycoprotein, regulates the complement system to prevent immune damage. In cancer, altered CD59 expression allows tumors to evade immune surveillance, promote growth, and resist certain immunotherapies. Targeting CD59 could enhance cancer treatment strategies by boosting the immune response against tumors. Herein, we present a one-step synthesis of Polyethyleneimine (PEI) functionalized graphene quantum dots (Lf-GQDs) from weathered lemon leaf extract. The fabricated Lf-GQDs were successfully used for the quantitative detection of the cluster of CD59 antigen that is reported for its expression in different types of cancer. In this work, we utilized orientation-based attachment of CD59 antibody (Anti-CD59). Our findings reveal that, instead of using random serial addition of antigen or antibody, oriented conjugation saves accumulated concentration offering greater sensitivity and selectivity. The Anti-CD59@Lf-GQDs immunosensor was fabricated using the oriented conjugation of antibodies onto the Lf-GQDs surface. Besides, the fabricated immunosensor demonstrated detection of CD59 in the range of 0.01 to 40.0 ng mL−1 with a low detection limit of 5.3 pg mL−1. Besides, the cellular uptake potential of the synthesized Lf-GQDs was also performed in A549 cells using a bioimaging study. The present approach represents the optimal utilization of Anti-CD59 and CD59 antigen. This approach could afford a pathway for constructing oriented conjugation of antibodies on the nanomaterials-based immunosensor for different biomarkers detection.

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Acknowledgements

The authors are grateful to the SERB-DST, Government of India (ECR/2017/000905) for research funding and HRPIPER, Shirpur, for providing the required facilities. We are thankful to STIC Cochin and IIT, Roorkee for providing characterization facilities.

Funding

The authors are grateful to the SERB-DST, Government of India (ECR/2017/000905) for research funding.

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Authors and Affiliations

  1. Department of Pharmaceutical Chemistry, H. R. Patel Institute of Pharmaceutical Education and Research, Shirpur - 425405, (MS), India

    Rahul Shankar Tade & Pravin Onkar Patil

  2. Department of Bioscience and Bioengineering, Indian Institute of Technology, Roorkee, 247667, Uttarakhand, India

    Ashish Kalkal

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  1. Rahul Shankar Tade
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Contributions

Pravin O. Patil: Conceptualization, review of original draft, supervision, editing, funding acquisition. Rahul S. Tade: Conceptualization, experimentation, data curation, writing of original draft, Ashish Kalkal: Review of original draft, comments and visualization.

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Correspondence to Pravin Onkar Patil.

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Tade, R.S., Kalkal, A. & Patil, P.O. Functionalized Graphene Quantum Dots (GQDs) based Label-Free Optical Fluorescence Sensor for CD59 Antigen Detection and Cellular Bioimaging. J Fluoresc (2023). https://doi.org/10.1007/s10895-023-03501-y

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