Method and system of microfluidic immunoassay using magnetic beads
Inventors
Barry, Andrew • Provencher, Laurel • Cohen, Seth • Chen, I-Jane • Yan, Jun • Wang, Jingjing
Assignees
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Publication Number
US-9835623-B2
Publication Date
2017-12-05
Expiration Date
Abstract
A microfluidic Western blot method and system including a microfluidic western blot method for immunoassay of proteins, the method including introducing a sample including the proteins onto a chip; electrophoretically separating the proteins; binding the separated proteins to beads to form protein-attached beads, the beads being magnetic; flowing the protein-attached beads into a magnetic holding region; applying a magnetic field to the magnetic holding region to fix the protein-attached beads in place within the magnetic holding region; binding primary antibodies to target proteins on the protein-attached beads; binding secondary antibodies to the bound primary antibodies; and detecting the bound secondary antibodies.
Core Innovation
The document describes a microfluidic western blot immunoassay in which proteins are electrophoretically separated on a microfluidic chip and then bound to magnetic beads to form protein-attached beads. The sample including the proteins is introduced onto the microfluidic chip, and the separated proteins are bound to magnetic beads within the microfluidic workflow.
The protein-attached beads are flowed into a magnetic holding region of the microfluidic chip, and a magnetic field is applied to fix the protein-attached beads in place. The magnetic field is applied before the binding of the primary antibodies, and primary antibodies are bound to target proteins on the protein-attached beads within the microfluidic chip.
Secondary antibodies are then bound to the bound primary antibodies within the microfluidic chip, and bound secondary antibodies are detected. The document further describes a microfluidic layout that includes a sample well, a separation region, a magnetic holding region, and additional regions such as a peak detection region, with detection carried out using an optical detector.
Claims Coverage
The partial content provides two independent claims. Across these claims, the inventive features focus on a microfluidic western blot immunoassay workflow that electrophoretically separates proteins, captures them on magnetic beads, fixes the bead complexes in a magnetic holding region using a magnetic field applied before primary antibody binding, then performs primary and secondary antibody binding followed by detection within the microfluidic chip.
Magnetic bead capture within a microfluidic western blot immunoassay workflow
A microfluidic western blot method for immunoassay of proteins introducing a sample including the proteins onto a microfluidic chip, electrophoretically separating the proteins, binding the separated proteins to beads to form protein-attached beads, the beads being magnetic, and detecting bound secondary antibodies within the microfluidic chip.
Magnetic holding region immobilization with magnetic field applied before primary antibody binding
Flowing the protein-attached beads into a magnetic holding region of the microfluidic chip and applying a magnetic field to the magnetic holding region to fix the protein-attached beads in place within the magnetic holding region, wherein applying of the magnetic field is before the binding of the primary antibodies; binding primary antibodies to target proteins on the protein-attached beads and binding secondary antibodies to the bound primary antibodies within the microfluidic chip.
Chip architecture linking sample well, separation region, and magnetic holding region
Providing a microfluidic chip having a substrate defining a sample well, a separation region operably coupled to the sample well, and a magnetic holding region operably coupled to the separation region; introducing the sample into the sample well; flowing the sample into the separation region; applying a voltage across the separation region to electrophoretically separate the proteins; binding the electrophoretically separated proteins to beads to form protein-attached beads, the beads being magnetic; flowing the protein-attached beads into the magnetic holding region; applying a magnetic field to fix the protein-attached beads in place; binding primary antibodies and binding secondary antibodies within the microfluidic chip; and detecting bound secondary antibodies within the microfluidic chip, with the magnetic field applied before the binding of the primary antibodies.
The independent claims are centered on electrophoretic separation on a microfluidic chip, capture on magnetic beads to form protein-attached beads, immobilization in a magnetic holding region by applying a magnetic field before primary antibody binding, followed by primary and secondary antibody binding and detection within the microfluidic chip.
Stated Advantages
Not explicitly described in patent.
Documented Applications
Not explicitly described in patent.
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