Automated 3D-printed unibody immunoarray for chemiluminescence detection of cancer biomarker proteins

Abstract

A low cost three-dimensional (3D) printed clear plastic microfluidic device was fabricated for fast, low cost automated protein detection. The unibody device features three reagent reservoirs, an efficient 3D network for passive mixing, and an optically transparent detection chamber housing a glass capture antibody array for measuring chemiluminescence output with a CCD camera. Sandwich type assays were built onto the glass arrays using a multi-labeled detection antibody-polyHRP (HRP = horseradish peroxidase). Total assay time was ~30 min in a complete automated assay employing a programmable syringe pump so that the protocol required minimal operator intervention. The device was used for multiplexed detection of prostate cancer biomarker proteins prostate specific antigen (PSA) and platelet factor 4 (PF-4). Detection limits of 0.5 pg mL−1 were achieved for these proteins in diluted serum with log dynamic ranges of four orders of magnitude. Good accuracy vs ELISA was validated by analyzing human serum samples. This prototype device holds good promise for further development as a point-of-care cancer diagnostics tool.

Disposable Sensors in Diagnostics, Food, and Environmental Monitoring

“…It is unfortunately not simple to implement every bioassay using paper alone; the use of paper is generally limited to applications compatible with open‐channel microfluidics. Materials, such as glass, PDMS, PS, and others, have also been used extensively in constructing disposable sensors. These materials can generally be transformed into functional, single‐use microfluidic POCT sensors through microfabrication or molding/replication methods and are used for on‐site detection of various analytes including antibiotics or other substances .…”

Advances in Reagents Storage and Release in Self‐Contained Point‐of‐Care Devices

“…CL-based analytical tools have already attracted great attentions because of their intrinsic merits, including the high sensitivity, high signal-to-noise ratio, and simple instruments. [37][38][39] H 2 O 2 -luminol is one of the commonly used CL substrates. However, H 2 O 2 -liminol needs to be stored separately at low temperature and mixed in situ, which may restrict its applications in POC areas.…”

3D-Printed Biosensor Arrays for Medical Diagnostics

“…The device comprised two main functions: mixing of the ATP sample with the luminescence reagent mixture (Luciferin/Luciferase mixture) and a detection chamber that brings the produced luminescence close to a silicon photomultiplier detector [ 93 ]. Tang et al utilized 3D printing to fabricate a unibody ELISA-inspired chemiluminescence assay to detect and quantify prostate specific antigen (PSA) and platelet factor-4 (PF-4) as cancer biomarker proteins ( Figure 6 C) [ 94 ]. They proposed a design that can reduce the assay time to 30 min while approaching an ultra-low sensitivity.…”

“…( C ) Unibody 3D-printed microfluidic chip for detection of PSA and PF-4. Reproduced with permission from [ 94 ]. Copyright (2017) Royal Society of Chemistry.…”