Antibody-Conjugated Magnetic Beads for Sperm Sexing Using a Multi-Wall Carbon Nanotube Microfluidic Device

Abstract

This study proposes a microfluidic device used for X-/Y-sperm separation based on monoclonal antibody-conjugated magnetic beads, which become positively charged in the flow system. Y-sperms were selectively captured via a monoclonal antibody and transferred onto the microfluidic device and were discarded, so that X-sperms can be isolated and commercially exploited for fertilization demands of female cattle in dairy industry. Therefore, the research team used monoclonal antibody-conjugated magnetic beads to increase the force that causes the Y-sperm to be pulled out of the system, leaving only the X-sperm for further use. The experimental design was divided into the following: Model 1, the microfluid system for sorting positive magnetic beads, which yielded 100% separation; Model 2, the sorting of monoclonal antibody-conjugated magnetic beads in the fluid system, yielding 98.84% microcirculation; Model 3, the sorting of monoclonal antibody-conjugated magnetic beads with sperm in the microfluid system, yielding 80.12% microcirculation. Moreover, the fabrication microfluidic system had thin film electrodes created via UV lithography and MWCNTs electrode structure capable of erecting an electrode wall 1500 µm above the floor with a flow channel width of only 100 µm. The system was tested using a constant flow rate of 2 µL/min and X-/Y-sperm were separated using carbon nanotube electrodes at 2.5 V. The structure created with the use of vertical electrodes and monoclonal antibody-conjugated magnetic beads technique produced a higher effective rejection effect and was able to remove a large number of unwanted sperm from the system with 80.12% efficiency.

Breaking the clean room barrier: exploring low-cost alternatives for microfluidic devices

“…Subsequently, a patterned substrate known as the “master” is created using expensive techniques, which offer excellent resolution. These techniques include photolithography (250 nm), nanoimprint lithography (15 nm), x-ray lithography (15 nm), or electron beam lithography (10 nm), but are primarily used in large-scale manufacturing facilities ( Maldonado & Peckerar, 2016 ; Gale et al, 2018 ; Thuau et al, 2018 ; Matsumoto et al, 2020 ; Phiphattanaphiphop et al, 2022 ). Consequently, there has been a growing interest in employing low-cost manufacturing techniques, including micromilling (25 μm), 3D printing (5–100 μm), and laser cutting (25 μm) ( Gale et al, 2018 ; Rusling, 2018 ; Hamilton et al, 2021 ; Šakalys et al, 2021 ; Preetam et al, 2022 ; Qin S et al, 2022 ).…”

Microfluidic Platforms for Real‐Time In Situ Monitoring of Biomarkers for Cellular Processes

“…[40] Multiwall carbon nanotubes have also been used in microfluidic platforms for sperm sexing, exhibiting 80.12% microcirculation. [41] Li et al reported a double-helix microfiber coupler coated with graphene oxide as a microfluidic platform, with a low detection limit and high sensitivity. [42] In recent years, several semiconductor nanomaterials, due to their excellent fluorescence properties, have been widely used to construct microfluidic fluorescence biosensors.…”

“…[ 40 ] Multiwall carbon nanotubes have also been used in microfluidic platforms for sperm sexing, exhibiting 80.12% microcirculation. [ 41 ] Li et al. reported a double‐helix microfiber coupler coated with graphene oxide as a microfluidic platform, with a low detection limit and high sensitivity.…”