Bis(sulfosuccinimidyl)suberate-Based Helix-Shaped Microchannels as Enhancers of Biomolecule Isolation from Liquid Biopsies

Abstract

Most studies of ultrasensitive diagnosis of biomolecules from liquid specimens are limited by problems during sample preparation steps, including enrichment and isolation of biomolecules. Here we report a novel platform combining bis­(sulfosuccinimidyl)­suberate (BS3) and helix-shaped microchannels (BSH) to change the sample preparation paradigm. This BSH system is composed of BS3 for pathogen enrichment and nucleic acid isolation by electrostatic and covalent interaction, and helix-shaped microchannels to minimize sample loss and remove bubbles in large liquid specimens without pH change. The system detected Mycobacterium tuberculosis following enrichment and isolation of 10 mL of liquefied sputum from 11 patients with tuberculosis. Moreover, the system identified KRAS mutations following cell-free DNA isolation of blood plasma from 10 patients with colorectal cancer. This system allows ultrasensitive diagnosis in various disease applications with large volumes of liquid samples.

Self-directed molecular diagnostics (SdMDx) system for COVID-19 via one-pot processing

“…Our previous studies have demonstrated that various biomaterial composites enable rapid, efficient, and affordable enrichment and separation of viral biomolecules for early clinical diagnosis [13] , [14] , [15] , [16] , [17] . Furthermore, our previous studies have shown that complex devices and expensive reagents are not needed for biomolecule separation steps, including pathogen enrichment and extraction [40] , [41] , [42] , [43] , [44] , [45] , [46] , and can be performed in POC settings. We developed a rapid and efficient SdMDx system with high sensitivity and specificity for the enrichment, amplification, and detection of SARS-CoV-2 and is self-operated and self-diagnosed at home as a POC test.…”

Gene-Based Diagnosis of Tuberculosis from Oral Swabs with a New Generation Pathogen Enrichment Technique

“…S1 depicts the overall workflow of the SLIM assay. The principle and the detailed structure of the SLIM assay have been described previously ( 9 , 10 , 13 , 37 ). Briefly, the SLIM assay is based on a combination of a microfluidic platform with low-cost thin film and homobifunctional imidoesters (HIs) reagents for MTB enrichment and DNA extraction from the oral swab samples.…”

Recent Advances in Device Engineering and Computational Analysis for Characterization of Cell-Released Cancer Biomarkers

“…In order to resolve concerns regarding loss of ctDNA, microfluidic-based techniques have been implemented for cfDNA isolation, in which the incorporation of microstructures, such as microbeads [ 84 ], micropillars [ 82 ], and microwells [ 85 , 86 ], enhances the cfDNA/ctDNA extraction performance and consequently shortens the processing time ( Figure 3 A,B). The integration of operations in the microfluidic chips has resulted in higher extraction performance.…”

“…( B ) In a novel microfluidic chip that contains helix-shaped microchambers to avoid forming bubbles, bis-(sulfosuccinimidyl)suberate reagent is used in lieu of chaotropic and pH modifier reagents to capture cfDNA while this device is capable of enriching pathogens in blood at the same time. Reproduced with permission from [ 86 ]. Copyright 2020 American Chemical Society.…”