Paper and Fiber-Based Bio-Diagnostic Platforms: Current Challenges and Future Needs

In this perspective article, some of the latest paper and fiber-based bio-analytical platforms are summarized, along with their fabrication strategies, the processing behind the product development, and the embedded systems in which paper or fiber materials were integrated. The article also reviews bio-recognition applications of paper/fiber-based devices, the detected analytes of interest, applied detection techniques, the related evaluation parameters, the type and duration of the assays, as well as the advantages and disadvantages of each technique. Moreover, some of the existing challenges of utilizing paper and/or fiber materials are discussed. These include control over the physical characteristics (porosity, permeability, wettability) and the chemical properties (surface functionality) of paper/fiber materials are discussed. Other aspects of the review focus on shelf life, the multi-functionality of the platforms, readout strategies, and other challenges that have to be addressed in order to obtain reliable detection outcomes. Keywords: paper-based bio-analytical devices; shelf life; equipment-free bio-recognition; flow rate; readout strategies; multi-functional platform

Design of a surface plasmon resonance immunoassay for therapeutic drug monitoring of amikacin

The therapeutic drug monitoring (TDM) of pharmaceutical drugs with narrow therapeutic ranges is of great importance in the clinical setting. It provides useful information towards the enhancement of drug therapies, aiding in dosage control and toxicity risk management. Amikacin is an aminoglycoside antibiotic commonly used in neonatal therapies that is indicated for TDM due to the toxicity risks inherent in its use. Current techniques for TDM such as high performance liquid chromatography (HPLC) and gas chromatography-mass spectrometry (GC-MS) are costly, time consuming, and cannot be performed at the site of action. Over the last decades, surface plasmon resonance (SPR) biosensors have become increasingly popular in clinical diagnostics due to their ability to detect biomolecular interactions in real-time. We present an SPR-based competitive immunoassay for the detection of the antibiotic amikacin, suitable for TDM in both adults and neonates. We have obtained high specificity and sensitivity levels with an IC value of 1.4 ng/mL and a limit of detection of 0.13 ng/mL, which comfortably comply with the drug's therapeutic range. Simple dilution of serum can therefore be sufficient to analyze low-volume real samples from neonates, increasing the potential of the methodology for TDM. Compared to current TDM conventional methods, this SPR-based immunoassay can provide advantages such as simplicity, potential portability, and label-free measurements with the possibility of high throughput. This work is the foundation towards the development of an integrated, simple use, highly sensitive, fast, and point-of-care sensing platform for the opportune TDM of antibiotics and other drugs in a clinical setting

Design of a surface plasmon resonance immunoassay for therapeutic drug monitoring of amikacin

The therapeutic drug monitoring (TDM) of pharmaceutical drugs with narrow therapeutic ranges is of great importance in the clinical setting. It provides useful information towards the enhancement of drug therapies, aiding in dosage control and toxicity risk management. Amikacin is an aminoglycoside antibiotic commonly used in neonatal therapies that is indicated for TDM due to the toxicity risks inherent in its use. Current techniques for TDM such as high performance liquid chromatography (HPLC) and gas chromatography-mass spectrometry (GC-MS) are costly, time consuming, and cannot be performed at the site of action. Over the last decades, surface plasmon resonance (SPR) biosensors have become increasingly popular in clinical diagnostics due to their ability to detect biomolecular interactions in real-time. We present an SPR-based competitive immunoassay for the detection of the antibiotic amikacin, suitable for TDM in both adults and neonates. We have obtained high specificity and sensitivity levels with an IC value of 1.4 ng/mL and a limit of detection of 0.13 ng/mL, which comfortably comply with the drug's therapeutic range. Simple dilution of serum can therefore be sufficient to analyze low-volume real samples from neonates, increasing the potential of the methodology for TDM. Compared to current TDM conventional methods, this SPR-based immunoassay can provide advantages such as simplicity, potential portability, and label-free measurements with the possibility of high throughput. This work is the foundation towards the development of an integrated, simple use, highly sensitive, fast, and point-of-care sensing platform for the opportune TDM of antibiotics and other drugs in a clinical setting

Mechanistic Analysis and Control of Electro‐Fabrication with Soft Matter: Polysaccharide Self‐Assembly with Electrochemically Produced Metal Ions

Abstract Electro‐fabrication is an emerging additive manufacturing approach that includes a variety of mechanisms and crosslinking chemistries to induce the self‐assembly‐ of soft matter. Within electro‐fabrication techniques, anodic electro‐fabrication is of great interest, because it can use the electrode as a sacrificial source of metal ions for the controlled formation of chelated hydrogels. However, it remains challenging to understand how the various solution conditions (e.g., buffer capacity, ligands) and imposed voltages must be controlled to achieve the chelation and sol‐gel transition. Herein, a comprehensive thermodynamics‐based framework is built to guide the selection of conditions for the self ‐assembly of polysaccharide hydrogels by crosslinking with transition metal ions generated via corrosion. This methodology is demonstrated through real‐time spectro‐electrochemical monitoring and employed for the electro‐fabrication of chitosan hydrogels chelated with Au3+, Ag+, Cu2+, and Pd2+, and alginate hydrogels chelated with Cu2+. The presented guidelines are intended to extend the capabilities of the controlled fabrication of assemblies crosslinked with metal ions, opening new horizons for unprecedented materials design with precisely tailored structures and functions

Polymethacrylate Sphere-Based Assay for Ultrasensitive miRNA Detection

Although microRNAs (miRNAs) have emerged as increasingly important target analytes, their biorecognition remains challenging due to their small size, high sequence homology, and low abundance in clinical samples. Nanospheres and microspheres have also gained increasing attention in biosensor applications due to their high specific surface area and the wide variety of compositions available. In this study, chemically designed and synthesized microspheres with active functional groups were used to promote effective miRNA immobilization resulting in better biorecognition. Upon conjugation with fluorescence-labeled complimentary probes, acylate-based spheres have indirectly detected MiR159, offering significantly enhanced analytical sensitivity, specificity, and accuracy while yielding a considerably low limit of detection (LOD) of 40 picomolar. Furthermore, MiR159 presence, which is known to be inversely correlated to breast cancer incidence and progression, was successfully detected in a competitive assay, which is promising for upgrading the current assay to clinical use

Polymethacrylate Coated Electrospun PHB Fibers as a Functionalized Platform for Bio-Diagnostics: Confirmation Analysis on the Presence of Immobilized IgG Antibodies against Dengue Virus.

In this article, a combination of far field electrospinning (FFES) and free-radical polymerization has been used to create a unique platform for protein immobilization via the physical attachment of biomolecules to the surface of the fiber mats. The large specific surface area of the fibers with its tailored chemistry provides a desirable platform for effective analyte-surface interaction. The detailed analysis of protein immobilization on a newly developed bio-receptive surface plays a vital role to gauge its advantages in bio-diagnostic applications. We relied on scanning electron microscopy (SEM), diameter range analysis, and X-ray photoelectron spectroscopy (XPS), along with thermal gravimetric analysis (TGA), water-in-air contact angle analysis (WCA), Fourier transform infrared spectroscopy (FTIR), and atomic force microscopy (AFM) to study our developed platforms and to provide valuable information regarding the presence of biomolecular entities on the surface. Detailed analyses of the fiber mats before and after antibody immobilization have shown obvious changes on the surface of the bioreceptive surface including: (i) an additional peak corresponding to the presence of an antibody in TGA analysis; (ii) extra FTIR peaks corresponding to the presence of antibodies on the coated fiber platforms; and (iii) a clear alteration in surface roughness recorded by AFM analysis. Confirmation analyses on protein immobilization are of great importance as they underlay substantial grounds for various biosensing applications

Rapid Lipid Content Screening in Neochloris oleoabundans Utilizing Carbon-Based Dielectrophoresis.

In this study, we carried out a heterogeneous cytoplasmic lipid content screening of Neochloris oleoabundans microalgae by dielectrophoresis (DEP), using castellated glassy carbon microelectrodes in a PDMS microchannel. For this purpose, microalgae were cultured in nitrogen-replete (N+) and nitrogen-deplete (N-) suspensions to promote low and high cytoplasmic lipid production in cells, respectively. Experiments were carried out over a wide frequency window (100 kHz-30 MHz) at a fixed amplitude of 7 VPP. The results showed a statistically significant difference between the dielectrophoretic behavior of N+ and N- cells at low frequencies (100-800 kHz), whereas a weak response was observed for mid- and high frequencies (1-30 MHz). Additionally, a finite element analysis using a 3D model was conducted to determine the dielectrophoretic trapping zones across the electrode gaps. These results suggest that low-cost glassy carbon is a reliable material for microalgae classification-between low and high cytoplasmic lipid content-through DEP, providing a fast and straightforward mechanism