Issue 1, 2024

Kelvin probe force microscopy on patterned large-area biofunctionalized surfaces: a reliable ultrasensitive platform for biomarker detection

Abstract

Kelvin probe force microscopy (KPFM) allows the detection of single binding events between immunoglobulins (IgM, IgG) and their cognate antibodies (anti-IgM, anti-IgG). Here an insight into the reliability and robustness of the methodology is provided. Our method is based on imaging the surface potential shift occurring on a dense layer of ∼5 × 107 antibodies physisorbed on a 50 μm × 90 μm area when assayed with increasing concentrations of antigens in phosphate buffer saline (PBS) standard solutions, in air and at a fixed scanning location. A comprehensive investigation of the influence of the main experimental parameters that may interfere with the outcomes of KPFM immune-assay is provided, showing the robustness and reliability of our approach. The data are supported also by a thorough polarization modulation infrared reflection–absorption spectroscopy (PM-IRRAS) analysis of the physisorbed biolayer, in the spectral region of the amide I, amide II and amide A bands. Our findings demonstrate that a 10 min incubation in 500 μL PBS encompassing ≈ 30 antigens (100 zM) triggers an extended surface potential shift that involves the whole investigated area. Such a shift quickly saturates at increasing ligand concentration, showing that the developed sensing platform works as an OFF/ON detector, capable of assessing the presence of a few specific biomarkers in a given assay volume. The reliability of the developed methodology KPFM is an important asset in single molecule detections at a wide electrode interface.

Graphical abstract: Kelvin probe force microscopy on patterned large-area biofunctionalized surfaces: a reliable ultrasensitive platform for biomarker detection

Supplementary files

Article information

Article type
Paper
Submitted
29 ago 2023
Accepted
18 nov 2023
First published
20 nov 2023
This article is Open Access
Creative Commons BY-NC license

J. Mater. Chem. C, 2024,12, 73-79

Kelvin probe force microscopy on patterned large-area biofunctionalized surfaces: a reliable ultrasensitive platform for biomarker detection

C. Di Franco, M. Piscitelli, E. Macchia, C. Scandurra, M. Catacchio, L. Torsi and G. Scamarcio, J. Mater. Chem. C, 2024, 12, 73 DOI: 10.1039/D3TC03110A

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