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Dissolving APTES in methanol with some water traces resulted in forming an APTES layer with 8.1 Å thickness based on spectroscopic ellipsometry measurements, which is close to the thickness of a monolayer . As previously mentioned, the amount of water in a solution and native -OH groups on an oxide substrate is critical in controlling the level of polymerization of silane molecules. XPS studies revealed that, compared to a deposition in toluene, the presence of N and C was three and two times greater, respectively, indicating more than one monolayer of APTES. Studies conducted by Pasternack et al. confirmed that lower APTES concentrations (0.1%) dissolved in toluene could create densely packed propyl chains on SiO2 when the reaction was carried out at 70 °C for less than 1 h. The deposition of APTES, in this case, was conducted on silicon wafers in a closed container (i.e., a toluene solution, and an APTES concentration range of 1–33%) backfilled with N2 gas.
Because of their morphologic versatility, chemical stability, and physicochemical interfacial properties, oxide materials have been used extensively in biosensor development . Oxide surfaces are surfaces that have hydroxyl groups (-OH) bearing high surface energy that can rapidly interact and form a covalent bond with silane molecules 12,13,14,15. The possibility for electrochemical modification of the surface functional groups of a self-assembled siloxane-anchored monolayer was explored. Aptamers are able to bind an ample range of non-nucleic acid targets with great specificity and affinity. The study finds that using toluene results in thicker APTES films ( Å) compared to PBS, which yields thinner films (8-13 Å). There was no increase in total protein surface density due to blocking with BSA for surfaces with already high protein coverage.
The amine group of the silane molecule is used to attach bioreceptors to the surface (Figure 1A); however, it can also form a weak hydrogen bond with the surface silanol group or with the silanol group of other silane molecules (Figure 1B). In addition, the silanol groups of different silane molecules can interact with each other, thereby forming polymeric structures (Figure 1B(5–7)). This procedure was repeated three times and 2% APTES in 100 ml of methanol was used to cover the NPs surfaces with silane molecules. They simply introduced 0.3 mL of APTES via an argon carrier gas (27 sccm) to a deposition chamber containing the oxide surfaces, which were hydroxylated with oxygen plasma.
Amino-functionalized organic films were prepared by self-assembling 3-aminopropyltriethoxysilane (APTES) on silicon wafers in either anhydrous toluene or phosphate-buffered saline (PBS) for varied deposition times. APTES films exhibit varying thicknesses and structural characteristics, with thicker films observed in anhydrous toluene (ranging from 10 Å to 144 Å) compared to thinner films produced in phosphate-buffered saline (8 Å to 13 Å). The investigation reveals that the formation and structure of APTES films on silicon substrates are significantly influenced by deposition conditions, including the choice of solvent and deposition time.
A condensation reaction can happen between two APTES molecules and/or between the oxide surface and an APTES molecule . In the absence of water or if the water content is too low, the silanol formation could be incomplete 10,44,52,53. The water content determines the number of hydrolyzed groups during the hydrolysis step in the APTES molecule (hydrolysis of one, two, or three ethoxy groups) . The quality of the silane surface, including its morphology, strongly depends on the initial hydrolysis step, making it very critical . (A) Schematic representation of an APTES molecule, activation, and reaction with the hydroxylated oxide surface in an ideal situation. APTES has three functional reactive ethoxy groups and one amine group per one silane molecule (Figure 1).
Amino-terminated organic layers were deposited onto silicon (Si) wafers and PS layers via 3-aminopropyltriethoxysilane (APTES) prepared in freshly hydrolysis solution. XPS results revealed the presence of Fe, O, C, and N, consistent with the development of interlinking silane molecules on a NP’s surface. Karade et al. conducted a ninhydrin colorimetric assay to monitor the monolayer formation through the quantification of the amine groups on the NPs . Additionally, APTES grafting on Fe2O3 magnetic NPs can prevent oxidative damage and play a role in the hydrophilicity or hydrophobicity of the NP surface, which is important for further drug conjugation through terminal amine groups. Moreover, the experimental grafting density was very close to the theoretically predicted APTES monolayer density. The study revealed that the equilibrium adsorption capacity was 301.2 mg/g for the highest grafting density during incubations at 70 °C using 2% APTES. The formation of more APTES monolayers on nanoparticles was proposed by Liu et al., where good quality silane layers were formed when the reaction was carried out at an elevated temperature using methanol/toluene mixed solvents .
Howarter et al. showed that prolonged incubation times of 24–74 h resulted in thicker APTES layer formation, confirmed by ellipsometry measurements ranging between a 5–16.3 nm thickness . Generally, after a sufficient cleaning and surface activation process, the oxide surface becomes hydrophilic with a water contact angle (WCA) ranging from ≤ 5 degrees to 20–30 degrees 70,75 and a surface roughness of 0.45 nm . For example, X-ray photoelectron spectroscopy (XPS) revealed an increased number of -OH groups on oxygen plasma-treated surfaces relative to chemically-treated surfaces . The formation of hydrogen bonds can decrease the number of silane molecules that are supposed to be covalently attached to the surface . In the phase separation step, the reaction medium loses its homogeneity to form more tangible polymeric structures .
Most NPs coated with APTES are used in applications such as for chemical sensors, cosmetic products (e.g., UV scattering effect in sun cream), drug delivery, magnetic resonance imaging, and bioimaging . Table 2 summarizes all the referenced solution-phase and vapor-phase processes and their monolayer characterization results. After deposition, the argon and APTES were removed, and the pressure was reduced to 75 × 10−7 Torr. Like YES CVD, there is some variation in the details of MLD processes, for example, they can be undertaken with or without high temperatures, a substrate dehydration, and nitrogen purges. This preceded the ten minute deposition of 500 L of APTES at 500 Torr, after which several nitrogen purges were used to remove excess silane. The desorption of APTES molecules from the surface was negligible in buffer solutions such as PBS compared to water.
The chemical reaction between the APTES and oxide nanoparticles’ (NP) surface is similar to the reaction with plain oxide surfaces. According to the molecular structure, an APTES monolayer should have an average chain length of ∼5–10 Å 5,44,64,65, a 0.5–0.8 nm thickness 44,53,66,67,68, an average density of 2.1–4.2 molecules per nm2 64,69 and a silanol group density of 5 per nm2 43,50. In an ideal situation, the attachment of the bioreceptors on an oxide surface via silane molecules should start with the hydrolysis of the silane molecules followed by condensation without the formation of additional bonds or different reaction routes.
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