The Role of Surface Chemistry in the Efficacy of Protein and DNA Microarrays for Label-Free Detection: An Overview

Chiodi, Elisa; Marn, Allison M.; Geib, Matthew T.; Ünlü, M. Selim

doi:10.3390/polym13071026

Cited by 17 publications

(9 citation statements)

References 128 publications

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“…A detailed table of other miniaturized pHcontrolled devices is also sown in SI. Electrochemical strategies, in addition to facilitate the electronic management, can ease the miniaturization of the control of chemical reactions using Silicon fabrication technologies that would allow a cost-effective production of chips to facilitate the access to the technology [11] . However, these attempts were not able to provide the necessary specifications to achieve the control of the chemistry that would allow them to reach wider use.…”

Section: Introductionmentioning

confidence: 99%

Universal control of proton concentration using an electrochemically generated acid compatible with miniaturization

2022

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Section: Introductionmentioning

confidence: 99%

Universal control of proton concentration using an electrochemically generated acid compatible with miniaturization

2022

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“…With a huge dynamic range in analyte size ranging from small molecules [5] to extracellular vesicles [49], this technology has enabled kinetic characterization of hundreds of different molecules. The combination of the IRIS platform with the surface chemistry provided by the MCP polymers has produced significant results in molecular characterization and single particle detection [10,[50][51][52][53] with minimal instrumentation requirements [54].…”

Section: Monolayer Polymeric Coatings: the Sweet Spotmentioning

confidence: 99%

The Effects of Three-Dimensional Ligand Immobilization on Kinetic Measurements in Biosensors

et al. 2022

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The field of biosensing is in constant evolution, propelled by the need for sensitive, reliable platforms that provide consistent results, especially in the drug development industry, where small molecule characterization is of uttermost relevance. Kinetic characterization of small biochemicals is particularly challenging, and has required sensor developers to find solutions to compensate for the lack of sensitivity of their instruments. In this regard, surface chemistry plays a crucial role. The ligands need to be efficiently immobilized on the sensor surface, and probe distribution, maintenance of their native structure and efficient diffusion of the analyte to the surface need to be optimized. In order to enhance the signal generated by low molecular weight targets, surface plasmon resonance sensors utilize a high density of probes on the surface by employing a thick dextran matrix, resulting in a three-dimensional, multilayer distribution of molecules. Despite increasing the binding signal, this method can generate artifacts, due to the diffusion dependence of surface binding, affecting the accuracy of measured affinity constants. On the other hand, when working with planar surface chemistries, an incredibly high sensitivity is required for low molecular weight analytes, and furthermore the standard method for immobilizing single layers of molecules based on self-assembled monolayers (SAM) of epoxysilane has been demonstrated to promote protein denaturation, thus being far from ideal. Here, we will give a concise overview of the impact of tridimensional immobilization of ligands on label-free biosensors, mostly focusing on the effect of diffusion on binding affinity constants measurements. We will comment on how multilayering of probes is certainly useful in terms of increasing the sensitivity of the sensor, but can cause steric hindrance, mass transport and other diffusion effects. On the other hand, probe monolayers on epoxysilane chemistries do not undergo diffusion effect but rather other artifacts can occur due to probe distortion. Finally, a combination of tridimensional polymeric chemistry and probe monolayer is presented and reviewed, showing advantages and disadvantages over the other two approaches.

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“…The literature reports countless approaches suited to surface modification purposes. In particular, polymeric coating is one of the most widely exploited techniques, since polymers extend into the third dimension, generating high cross-sectional densities of functional groups homogeneously distributed onto the surface, therefore resulting in an increased binding capacity of probes [ 10 , 11 ]. Moreover, polymeric coating offers several other advantages, such as the facility in tailoring their composition to adapt the coating to specific applications or material, high accessibility of the ligand for the interaction with the partner, lower degree of non-specific adsorption (meaning reduced background signal) and exceptional retention of the ligand 3D structure, all of which consequently improve a probe’s biological activity.…”

Section: Introductionmentioning

confidence: 99%

Optimization of Functional Group Concentration of N, N-Dimethylacrylamide-based Polymeric Coatings and Probe Immobilization for DNA and Protein Microarray Applications

et al. 2023

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We report here a deep investigation into the effect of the concentration of a polymeric coating’s functional groups on probe density immobilization with the aim of establishing the optimal formulation to be implemented in specific microarray applications. It is widely known that the ideal performance of a microarray strictly depends on the way probes are tethered to the surface since it influences the way they interact with the complementary target. The N, N-dimethylacrylamide-based polymeric coating introduced by our research group in 2004 has already proven to offer great flexibility for the customization of surface properties; here, we demonstrate that it also represents the perfect scaffold for the modulation of probe grafting. With this aim in mind, polymers with increasing concentrations of N-acryloyloxysuccinimide (NAS) were synthesized and the coating procedure optimized accordingly. These were then tested not only in DNA microarray assays, but also using protein probes (with different MWs) to establish which formulation improves the assay performance in specific applications. The flexibility of this polymeric platform allowed us also to investigate a different immobilization chemistry—specifically, click chemistry reactions, thanks to the insertion of azide groups into the polymer chains—and to evaluate possible differences generated by this modification.

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The Role of Surface Chemistry in the Efficacy of Protein and DNA Microarrays for Label-Free Detection: An Overview

Cited by 17 publications

References 128 publications

Universal control of proton concentration using an electrochemically generated acid compatible with miniaturization

Universal control of proton concentration using an electrochemically generated acid compatible with miniaturization

The Effects of Three-Dimensional Ligand Immobilization on Kinetic Measurements in Biosensors

Optimization of Functional Group Concentration of N, N-Dimethylacrylamide-based Polymeric Coatings and Probe Immobilization for DNA and Protein Microarray Applications

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