MSA: scalable DNA methylation screening BeadChip for high-throughput trait association studies.

The Infinium DNA Methylation BeadChips have significantly contributed to population-scale epigenetics research by enabling epigenome-wide trait association discoveries. Here, we design, describe, and experimentally verify a new iteration of this technology, the Methylation Screening Array (MSA), to focus on human trait screening and discovery. This array utilizes extensive data from previous Infinium platform-based epigenome-wide association studies (EWAS). It incorporates knowledge from the latest single-cell and cell type-resolution whole genome methylome profiles. The MSA is engineered to achieve scalable screening of epigenetics-trait association in an ultra-high sample throughput. Our design encompassed diverse human trait associations, including those with genetic, cellular, environmental, and demographical variables and human diseases such as genetic, neurodegenerative, cardiovascular, infectious, and immune diseases. We comprehensively evaluated this array's reproducibility, accuracy, and capacity for cell-type deconvolution and supporting 5-hydroxymethylation profiling in diverse human tissues. Our first atlas data using this platform uncovered the complex chromatin and tissue contexts of DNA modification variations and genetic variants linked to human phenotypes.

Recent advances of PLGA micro/nanoparticles for the delivery of biomacromolecular therapeutics.

Recent advancements in biopharmaceutical industry have facilitated the development of novel bioactive macromolecular therapeutics. One of the challenges towards the clinical use of these biomacromolecules lies in the selection of appropriate carriers to protect, deliver and release them in vivo to maximize their pharmacological effects. Micro/nanoparticles made from biodegradable poly (d,l-lactic-co-glycolic acid) (PLGA) have been explored as delivery vehicles for therapeutics. Due to their excellent biocompatibility and controllable biodegradability, PLGA micro/nanoparticles could protect macromolecules from instant degradation in vivo while allowing tunable release rate and profile. In this review, recent progress in the design, fabrication/formulation and application of PLGA based micro/nanoparticles for the controlled delivery of biomacromolecules are discussed. Special focuses will be on the novel loading methods and releasing mechanisms of macromolecules as well as the in vivo applications of therapeutic macromolecule-loaded PLGA micro/nanoparticles.

White Light-Emitting Multistimuli-Responsive Hydrogels with Lanthanides and Carbon Dots.

Polymers that confer changes in optical properties in response to chemical or mechanical cues offer diverse sensing applications, particularly if this stimuli response is accessible in humid or aqueous environments. In this study, luminescent hydrogels were fabricated using a facile aqueous process by incorporating lanthanide ions and carbon dots (CD) into a network of polyacrylamide and poly(acrylic acid). White luminescence was obtained by tuning the balance of blue-light-emitting CD to green- and red-light-emitting lanthanide ions. Exploiting the combined specific sensitivities of the different emitters, the luminescent hydrogel showed chromic responsiveness to multiple stimuli, including pH, organic vapors, transition-metal ions, and temperature. The white-light-emitting hydrogel was also stretchable with a fracture strain of 400%. We envision this photoluminescent hydrogel to be a versatile and multifunctional material for chemical and environmental sensing.

PEG-based autonomous capillary system with integrated microbead array for immunoassay.

Surface-tension-driven capillary systems (CSs) enable self-powered delivery of samples and reagents for bioassays and thus are especially suitable for point-of-care applications. Current silicon and polymer based CSs require extensive work in professional cleanroom for the fabrication of either the silicon device itself or the micromold for polymer processing. In this work, we fabricated a PEG-based CS in a one-step photopolymerization process without the requirement of any cleanroom work. Water, buffer and serum can flow autonomously in this CS and the liquid flow rate can be tuned by modification with surfactant solution of different concentrations. We further integrated an antibody-coated microbead array into this CS for the autonomous immunoassay of two tumor markers, prostate specific antigen (PSA) and human chorionic gonadotropin (hCG), in serum samples with a total assay time of less than 10min. The detection limits for the two tumor markers were at sub-nanogram per milliliter range which is lower than their clinical threshold concentrations for cancer diagnosis. Moreover, simultaneous and multiplex detection of the two tumor markers was also achieved by using spatially encoded microbeads. This low cost and easy-fabricated CS enables fast, multiplex and autonomous immunoassay for protein markers and has the potential to be applied for point-of-care diagnostics on real clinical samples.

Polyethylene glycol (PEG) gel arrays for differentiating oligopeptide fragments and on-chip protease assays.

Polyethylene glycol (PEG) hydrogel is permeable to biomolecules, but its permeability depends on the molecular weight of monomers and the concentration of monomer solutions. In this study, we show that PEG hydrogel made from 20% to 30% of PEG700 monomer is permeable to amino acids yet impermeable to oligopeptides. Because of its unique permeability, the gel can be used to detect protease by separating amino acids from oligopeptides when proteases cleave the oligopeptides and release amino acids. Based on this principle, an UV crosslinked gel array is fabricated on a chip for simultaneous detection of protease in up to 40 samples with only 1 µl of volume required for each sample. As a proof of concept, the on-chip protease assays are used to detect trypsin in buffer and serum. The detection limits are 1.2 nM in buffer and 17.7 nM in serum, which are comparable to conventional protease assays. Moreover, because only 1 µl of liquid is required, as little as 1.2 fmol of trypsin can be detected by using the on-chip assay. The protease assay also shows good specificity for trypsin and chymotrypsin. The gel array chip could be a useful miniaturized platform for high-throughput detection of different proteases and screening of their inhibitors.

Microfluidic immunoassay with plug-in liquid crystal for optical detection of antibody.

Recent advance in liquid crystal (LqC) based immunoassays enables label-free detection of antibody, but manual preparation of LqC cells and injection of LqC are required. In this work, we developed a new format of LqC-based immunoassay which is hosted in a microfluidic device. In this format, the orientations of LqC are strongly influenced by four channel walls surrounding the LqC. When the aspect ratio (depth/width) of the channel is smaller than 0.38, LqC orients homeotropically inside the microchannel and appears dark. After antigens bind to immobilized antibodies on the channel walls, a shift of the LqC appearance from dark to bright (due to the disruption of LqC orientation) can be visualized directly. To streamline the immunoassay process, a tubing cartridge loaded with a sample solution, washing buffers and a plug of LqC is connected to the microfluidic device. By using pressure-driven flow, the cartridge allows antigen/antibody binding, washing and optical detection to be accomplished in a sequential order. We demonstrate that this microfluidic immunoassay is able to detect anti-rabbit IgG with a naked-eye detection limit down to 1 µg mL(-1). This new format of immunoassay provides a simple and robust approach to perform LqC-based label-free immunodetection in microfluidic devices.

Gel pad array chip for high throughput and multi-analyte microbead-based immunoassays.

We present here a gel pad array chip for high-throughput and multi-analyte microbead-based immunoassays. The chip is fabricated by photo-patterning of two polymeric gels, polyacrylamide gel and polyethylene glycol (PEG) gel, on a glass slide. The resulting chip consists of 40 polyacrylamide gel pad array units for the immobilization of microbeads and each gel pad array is surrounded with a PEG micropillar ring to confine the samples within the microarray. As a proof of concept, this chip was tested for quantitative immunoassays for two model cancer markers, human chorionic gonadotropin (hCG) and prostate specific antigen (PSA), in serum samples. Detection limits below the physiological threshold level for cancer diagnosis were achieved with good inter- and intra-chip reproducibility. Moreover, by using spatial encoded microbeads, simultaneous detection of both hCG and PSA on each gel pad array is achieved with single filter fluorescence imaging. This gel pad array chip is easy to use, easy to fabricate with low cost materials and minimal equipment and reusable. It could be a useful tool for common biolabs to customize their own microbead array for multi-analyte immunoassays.

Multiplex detection platform for tumor markers and glucose in serum based on a microfluidic microparticle array.

We present a multiplex detection platform based on a microfluidic microparticle array to detect proteins and glucose in serum simultaneously. Multiplex detection of proteins and glucose was performed using biofunctionalized microparticles arrayed on gel-based microstructures integrated in microfluidics. The microparticles immobilized on these microstructures showed high stability under microfluidic flow conditions. With arrays of antibody-coated microbeads, microfluidic quantitative immunoassays for two protein tumor markers, human chorionic gonadotropin (hCG) and prostate specific antigen (PSA) were performed in serum samples with detection limits bellow the cut-off values for cancer diagnosis. Parallel to the immunoassays, quantitative enzymatic assays for glucose in the physiological concentration range were performed. Multiplex detection was achieved by using a spatially encoded microarray. By patterning antibody-coated microbeads and enzyme-containing microparticles on a novel mixed structure array, we successfully demonstrated simultaneous immunoassays (binding based assay) for proteins and an enzymatic assay (reaction kinetic based assay) for glucose. Our microparticle arrays could be potentially used for the detection of multiple categories of biomolecules (proteins, small metabolites and DNA) for clinical diagnostics and other biological applications.

Utilizing microfluidics to synthesize polyethylene glycol microbeads for Förster resonance energy transfer based glucose sensing.

Here, we utilize microfluidic droplet technology to generate photopolymerizeable polyethylene glycol (PEG) hydrogel microbeads incorporating a fluorescence-based glucose bioassay. A microfluidic T-junction and multiphase flow of fluorescein isothiocyanate dextran, tetramethyl rhodamine isothiocyanate concanavalin A, and PEG in water were used to generate microdroplets in a continuous stream of hexadecane. The microdroplets were photopolymerized mid-stream with ultraviolet light exposure to form PEG microbeads and were collected at the outlet for further analysis. Devices were prototyped in PDMS and generated highly monodisperse 72 ± 2 µm sized microbeads (measured after transfer into aqueous phase) at a continuous flow rate between 0.04 ml/h-0.06 ml/h. Scanning electron microscopy analysis was conducted to analyze and confirm microbead integrity and surface morphology. Glucose sensing was carried out using a Förster resonance energy transfer (FRET) based assay. A proportional fluorescence intensity increase was measured within a 1-10 mM glucose concentration range. Microfluidically synthesized microbeads encapsulating sensing biomolecules offer a quick and low cost method to generate monodisperse biosensors for a variety of applications including cell cultures systems, tissue engineering, etc.