3D Printed Microfluidic Devices for Solid-Phase Extraction and On-Chip Fluorescent Labeling of Preterm Birth Risk Biomarkers

Abstract

Solid-phase extraction (SPE) is a general preconcentration method for sample preparation that can be performed on a variety of specimens. The miniaturization of SPE within a 3D printed microfluidic device further allows for fast and simple extraction of analytes, while also enabling integration of SPE with other sample preparation and separation methods. Here, we present the development and application of a reversed-phase lauryl methacrylate-based monolith, formed in 3D printed microfluidic devices, which can selectively retain peptides and proteins. The effectiveness of these SPE monoliths and 3D printed microfluidic devices was tested using a panel of nine preterm birth biomarkers of varying hydrophobicities and ranging in mass from 2-470 kDa. The biomarkers were selectively retained, fluorescently labeled, and eluted separately from the excess fluorescent label in 3D printed microfluidic systems. These are the first results demonstrating microfluidic analysis processes on a complete panel of preterm birth biomarkers, an important step toward developing a miniaturized, fully integrated analysis system.

High-Resolution 3D Printing Fabrication of a Microfluidic Platform for Blood Plasma Separation

“…Devices 1-15 were printed with a hydrophilic resin containing NPS (resin A, see Section 2.1), which gave them a yellowish appearance. This effect was already observed in previous research, which used the same resin formulation [22,23]. For Device 16, NPS was replaced by Avo (resin B) which, unlike resin A, gave the devices a transparent appearance, facilitating the subsequent colorimetric analysis of the separated plasma when performed in situ in the device.…”

Immunoaffinity monoliths for multiplexed extraction of preterm birth biomarkers from human blood serum in 3D printed microfluidic devices

“…The microfluidic devices (Fig. 1A) were adapted from Bickham et al 19 and designed to have five separated channels, each with a 50 μm × 45 μm cross section. Each channel had a reservoir on one side of the device and a port on the other side for PTFE tubing (0.22 in ID × 0.042 in OD; Cole Parmer, Vernon Hills, IL) to connect to vacuum to flow analyte through the channel.…”

“…16,17 3D printing of microfluidic devices can overcome challenges associated with traditional fabrication techniques, and can enable novel designs that are otherwise inaccessible. 12,18–21 These 3D printed microfluidic devices can be used to facilitate sample preparation and biomarker analysis. For example, Bickham et al 19 used solid phase extraction monoliths to concentrate and label a panel of nine PTB biomarkers.…”

“…12,18–21 These 3D printed microfluidic devices can be used to facilitate sample preparation and biomarker analysis. For example, Bickham et al 19 used solid phase extraction monoliths to concentrate and label a panel of nine PTB biomarkers. Although this is a good method for enriching and labeling samples, PTB biomarkers must first be purified from blood serum, which cannot be performed on this type of monolith.…”

Emerging Lab-on-a-Chip Approaches for Liquid Biopsy in Lung Cancer: Status in CTCs and ctDNA Research and Clinical Validation

“…Nonetheless, the field has evolved rapidly with state-of-the-art technologies being introduced regularly. Problems such as air bubbles formation, leaking or precise flow controls that caused constraints in first generation microfluidics devices are not relevant at present, with cutting edge strategies of intelligent lab-on-a-chips incorporating automation and digitalization to on-chip single-cell analysis, miRNA detection and nucleic acids quantification and analysis [ 111 , 121 , 122 , 123 , 124 ]. However, the relatively new application of microfluidics technology for liquid biopsy translates into few clinical validation studies and trials involving microfluidic chips, which are especially scarce in the context of LCa.…”