Microfluidic apparatus and methods of use thereof
Apparatuses and methods are described herein for processing polynucleotides in a sealed path environment. The apparatuses include optical sensors to monitor operations and to track material usage for good manufacturing practice.
1. A microfluidic apparatus for forming a therapeutic polynucleotide, the apparatus comprising:
a seating mount for removably holding a microfluidic path device;
a plurality of pressure lines;
a plurality of fluid vials, wherein each fluid vial either comprises a fluidic line or is configured to couple with the fluidic line,
wherein each fluidic line and at least a subset of the pressure lines are configured to be biased against the microfluidic path device held in the seating mount to form a closed fluid path;
an optical sensor drive configured to move one or more of a plurality of optical sensors around the seating mount or around the plurality of vials; and
a controller comprising one or more processors, the one or more processors programmed to control the application of pressure through the pressure lines to drive fluidic movement in the microfluidic path device when the microfluidic path device is held in the seating mount, wherein the controller is configured to direct the synthesis of a synthetic template, direct an in vitro transcription (IVT) reaction using the template to form a therapeutic polynucleotide, and direct purification of the therapeutic polynucleotide in one or more microfluidic path devices held in the seating mount.
2. A microfluidic apparatus for forming a therapeutic polynucleotide, the apparatus comprising:
a seating mount for removably holding a microfluidic path device;
a plurality of pressure lines;
a plurality of fluid vials, wherein each fluid vial either comprises a fluidic line or is configured to couple with the fluidic line,
wherein each fluidic line and at least a subset of the pressure lines are configured to be biased against the microfluidic path device held in the seating mount to form a closed fluid path;
an optical sensor drive configured to move one or more of a plurality of optical sensors around the seating mount or around the plurality of vials; and
a controller comprising one or more processors, the one or more processors programmed to control the application of pressure through the pressure lines to drive fluidic movement in the microfluidic path device when the microfluidic path device is held in the seating mount, wherein the controller is configured to determine the contents of the fluid vials, transfer sub-microliter amounts of material from the fluid vials to one or more reactors in the microfluidic path device held in the seating mount, direct the synthesis of a synthetic template, direct an in vitro transcription (IVT) reaction using the template to form a therapeutic polynucleotide, and direct purification of the therapeutic polynucleotide in one or more microfluidic path devices held in the seating mount.
3. The apparatus of claim 1 , further comprising a reagent storage frame holding the plurality of fluid vials.
4. The apparatus of claim 1 , further comprising a plurality of optical sensors arranged to monitor fluid levels within the plurality of fluid vials and to monitor fluidic movement in the microfluidic path device when the microfluidic path device is held in the seating mount.
5. The apparatus of claim 1 , further comprising a fluid interface assembly configured to hold the fluidic lines.
6. The apparatus of claim 1 , further comprising a fluid interface assembly release control configured to release the fluid interface assembly from the apparatus.
7. The apparatus of claim 1 , further comprising a reagent storage frame release control configured to release the reagent storage frame from the apparatus.
8. The apparatus of claim 1 , further comprising a thermal control configured to modulate the temperature of at least one region of the microfluidic path device when the microfluidic path device is held in the seating mount.
9. The apparatus of claim 1 , further comprising a magnetic field applicator configured to apply a magnetic field to at least one region of the microfluidic path device when the microfluidic path device is seated in the seating mount.
10. The apparatus of claim 9 , wherein the magnetic field applicator comprises a control arm mounted to the reagent storage frame.
11. The apparatus of claim 1 , wherein the controller is configured to detect an identifying code on the fluid vial.
12. The apparatus of claim 1 , wherein the controller is configured to determine a level of a reagent held by one or more of the plurality of fluid vials.
13. The apparatus of claim 1 , further comprising one or more alignment pins configured to align the microfluidic path device in the seating mount.
14. The apparatus of claim 1 , further comprising an enclosure having a cover, configured to maintain the sterility of the apparatus during operation of the apparatus.
15. The apparatus of claim 1 , further comprising the microfluidic path device.
16. The apparatus of claim 1 , further comprising a fluid interface assembly configured to couple to the seating mount, the fluid interface assembly having a central opening through which the microfluidic path device may be imaged by one or more of a plurality of optical sensors, further wherein a distal end of each of the fluidic lines and pressure lines are arranged around a periphery of the central opening and configured to be sealed against the microfluidic device path held in the seating mount.
17. The apparatus of claim 1 , further comprising a signal detector configured to detect a signal from within the microfluidic path device.
18. The apparatus of claim 17 , wherein the signal is one or more of: a visible, fluorescent, UV absorbance or IR absorbance signal.
19. The apparatus of claim 17 , wherein the signal detector is configured to measure a nanoparticle size distribution.
20. A microfluidic apparatus for forming a therapeutic polynucleotide, the apparatus comprising:
a seating mount for removably holding a microfluidic path device;
a plurality of pressure lines;
a plurality of fluid vials, wherein each fluid vial either comprises a fluidic line or is configured to couple with the fluidic line,
wherein each fluidic line and at least a subset of the pressure lines are configured to be biased against the microfluidic path device held in the seating mount to form a closed fluid path;
a fluid interface assembly configured to couple to the seating mount, the fluid interface assembly having a central opening through which the microfluidic path device may be imaged by one or more of a plurality of optical sensors, further wherein a distal end of each of the fluidic lines and pressure lines are arranged around a periphery of the central opening and configured to be sealed against the microfluidic device path held in the seating mount; and
a controller comprising one or more processors, the one or more processors programmed to control the application of pressure through the pressure lines to drive fluidic movement in the microfluidic path device when the microfluidic path device is held in the seating mount, wherein the controller is configured to direct the synthesis of a synthetic template, direct an in vitro transcription (IVT) reaction using the template to form a therapeutic polynucleotide, and direct purification of the therapeutic polynucleotide in one or more microfluidic path devices held in the seating mount.
21. The apparatus of claim 20 , further comprising a reagent storage frame holding the plurality of fluid vials.
22. The apparatus of claim 20 , further comprising a plurality of optical sensors arranged to monitor fluid levels within the plurality of fluid vials and to monitor fluidic movement in the microfluidic path device when the microfluidic path device is held in the seating mount.
23. The apparatus of claim 20 , further comprising a fluid interface assembly configured to hold the fluidic lines.
24. The apparatus of claim 20 , further comprising a fluid interface assembly release control configured to release the fluid interface assembly from the apparatus.
25. The apparatus of claim 20 , further comprising a reagent storage frame release control configured to release the reagent storage frame from the apparatus.
26. The apparatus of claim 20 , further comprising a thermal control configured to modulate the temperature of at least one region of the microfluidic path device when the microfluidic path device is held in the seating mount.
27. The apparatus of claim 20 , further comprising a magnetic field applicator configured to apply a magnetic field to at least one region of the microfluidic path device when the microfluidic path device is seated in the seating mount.
28. The apparatus of claim 27 , wherein the magnetic field applicator comprises a control arm mounted to the reagent storage frame.
29. The apparatus of claim 20 , wherein the controller is configured to detect an identifying code on the fluid vial.
30. The apparatus of claim 20 , wherein the controller is configured to determine a level of a reagent held by one or more of the plurality of fluid vials.
31. The apparatus of claim 20 , further comprising an optical sensor drive configured to move one or more of a plurality of optical sensors around the seating mount or around the plurality of vials.
32. The apparatus of claim 20 , further comprising one or more alignment pins configured to align the microfluidic path device in the seating mount.
33. The apparatus of claim 20 , further comprising an enclosure having a cover, configured to maintain the sterility of the apparatus during operation of the apparatus.
34. The apparatus of claim 20 , further comprising the microfluidic path device.
35. The apparatus of claim 20 , further comprising a signal detector configured to detect a signal from within the microfluidic path device.
36. The apparatus of claim 35 , wherein the signal is one or more of: a visible, fluorescent, UV absorbance or IR absorbance signal.
37. The apparatus of claim 35 , wherein the signal detector is configured to measure a nanoparticle size distribution.
38. A microfluidic apparatus for forming a therapeutic polynucleotide, the apparatus comprising:
a seating mount for removably holding a microfluidic path device;
a plurality of pressure lines;
a plurality of fluid vials, wherein each fluid vial either comprises a fluidic line or is configured to couple with the fluidic line,
wherein each fluidic line and at least a subset of the pressure lines are configured to be biased against the microfluidic path device held in the seating mount to form a closed fluid path;
a fluid interface assembly configured to couple to the seating mount, the fluid interface assembly having a central opening through which the microfluidic path device may be imaged by one or more of a plurality of optical sensors, further wherein a distal end of each of the fluidic lines and pressure lines are arranged around a periphery of the central opening and configured to be sealed against the microfluidic device path held in the seating mount; and
a controller comprising one or more processors, the one or more processors programmed to control the application of pressure through the pressure lines to drive fluidic movement in the microfluidic path device when the microfluidic path device is held in the seating mount, wherein the controller is configured to determine the contents of the fluid vials, transfer sub-microliter amounts of material from the fluid vials to one or more reactors in the microfluidic path device held in the seating mount, direct the synthesis of a synthetic template, direct an in vitro transcription (IVT) reaction using the template to form a therapeutic polynucleotide, and direct purification of the therapeutic polynucleotide in one or more microfluidic path devices held in the seating mount.