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Cited by 38Photonic and Optical DevicesAdvanced Biosensing Techniques and ApplicationsPolymer Surface Interaction Studies

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Novel Rapid Protein Coating Technique for Silicon Photonic Biosensor to Improve Surface Morphology and Increase Bioreceptor Density

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Udomsom, Suruk, Mankong, Ukrit, Paengnakorn, Pathinan, Theera-Umpon, Nipon·2021·Coatings
DOI 10.3390/coatings11050595
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Abstract

Silicon photonic devices with either silicon or silicon nitride waveguides have increasingly been used in many applications besides communications, especially as sensors in label-free biosensing, where guided light signals are affected by biorecognition molecules immobilized on the surface. The coating of protein (i.e., bioreceptors) by biochemical process on the waveguide surface is a crucial step in creating a functionalized device that can be used for biosensing. As a conventional method that uses 3-aminopropryltriethoxysilane (APTES) and glutaraldehyde (GA), the APTES-GA method has the limitation of using a GA crosslink, of which the two functional groups can bind to nonspecific proteins, causing irregular binding. In this study, we proposed a new coating technique to avoid such problem by applying APTES silanization with 1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimide (EDC)-N-hydroxysuccinimide (NHS) protein crosslink, denoted by the APTES-(EDC/NHS) method. The EDC/NHS reaction was shown to be able to immobilize protein in ordered orientation due to consistent arrangement between a carboxylic group of protein molecules and an amine group of covalent-linked APTES on surface. By applying APTES silanization, we circumvented the use of hazardous cleaning agent in the conventional EDC/NHS technique. Several surface characterization techniques were carried out to assess and compare the two biocoating techniques, including scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), spectroscopic ellipsometry (SE), and atomic force microscopy (AFM). On silicon, the results of antihuman TNF-alpha antibody coating showed that the proposed APTES-(EDC/NHS) technique has better repeatability in terms of less roughness of the coated protein at 1.5 nm compared with 6.3 nm, due to the ordered arrangement of coated antibody molecules. On a silicon nitride waveguide device, the proposed APTES-(EDC/NHS) technique exhibits dense antibody immobilization on a waveguide in SEM images due to stable amide bond formation via EDC/NHS crosslink mechanism. The specificity of the immobilized antibodies was confirmed by enzyme-linked immunosorbent assays (ELISA), with an average optical density at 450 nm of 0.175 ± 0.01 compared with 0.064 ± 0.009 of negative control. The proposed technique also reduced the overall process time since proteins are crosslinked to the silanized waveguide surface in a single step.

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Metadata

Authors
Udomsom, S., Mankong, U., Paengnakorn, P., & Theera-Umpon, N.
Affiliations
  • Chiang Mai University (TH)
  • Biomedical Engineering Institute, Chiang Mai University, Chiang Mai 50200, Thailand
Corresponding author
  • Udomsom, Suruk
  • Mankong, Ukrit
ORCID
Venue
Coatings
Publisher
MDPI AG
Published
2021-05-18
Volume
11
Issue
5
Pages
595
ISSN
2079-6412 (ISSN-L)
DOI
10.3390/coatings11050595
OpenAlex
W3162329604
Cited by
38
References
62
Citations by year
2026: 5 · 2025: 15 · 2024: 7 · 2023: 5 · 2022: 6
FWCI
1.32
Type
journal article
Language
en
Open access
Licence
Locations
  • Coatings (publishedVersion, cc-by)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
Funders
Capacity Building Program for New Researcher 2018 from National Research Council of Thai-land (NRCT) (24834)
Topics
  • Physical Sciences › Engineering › Electrical and Electronic Engineering › Photonic and Optical Devices
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Advanced Biosensing Techniques and Applications
  • Physical Sciences › Materials Science › Surfaces, Coatings and Films › Polymer Surface Interaction Studies
Keywords
protein immobilization, APTES silanization, EDC/NHS coupling, silicon waveguides, silicon nitride waveguides, label-free biosensing, antibody immobilization, surface morphology, TNF-alpha antibodies, surface roughness, amide synthesis, glutaraldehyde crosslinking, surface functionalization, optical biosensors, surface plasmon resonance biosensor

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