Protein crystallization in microfluidic structures

• US 6,409,832 B2
• Filed: 03/30/2001
• Issued: 06/25/2002
• Est. Priority Date: 03/31/2000
• Status: Expired due to Term

• Current Assignee: PerkinElmer Health Sciences, Inc. (Revvity, Inc.)
• Original Assignee: Micronics Inc. (Sony Group Corp.)
• Inventors: Sygusch, Jurgen, Weigl, Bernhard H.
• Primary Examiner(s): HITESHEW, FELISA CARLA
• Application Number:
  US09/822,595
• Publication Number:
  US 20010027745A1
• Time in Patent Office: 452 Days
• Field of Search: 117/200, 117/206, 117/900, 422/245.1
• US Class Current: 117/206
• CPC Class Codes: B01F 33/30 Micromixers
• CPC Class Codes: B01F 33/3039 with mixing achieved by diffusion between layers
• CPC Class Codes: B01L 2200/0636 Focussing flows, e.g. to laminate flows
• CPC Class Codes: B01L 2200/0684 Venting, avoiding backpressure, avoid gas bubbles
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0867 Multiple inlets and one sample wells, e.g. mixing, dilution
• CPC Class Codes: B01L 2400/0481 squeezing of channels or chambers
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/0605 check valves
• CPC Class Codes: B01L 3/06 Crystallising dishes
• CPC Class Codes: B01L 3/5025 for parallel transport of multiple samples
• CPC Class Codes: B01L 3/50273 characterised by the means or forces applied to move the fluids micropumps F04B19/006, of the membrane type F04B43/043
• CPC Class Codes: B01L 3/502776 specially adapted for focusing or laminating flows
• CPC Class Codes: B01L 7/54 using spatial temperature gradients
• CPC Class Codes: C30B 27/00 Single-crystal growth under a protective fluid
• CPC Class Codes: C30B 29/58 Macromolecular compounds
• CPC Class Codes: C30B 30/08 in conditions of zero-gravity or low gravity
• CPC Class Codes: C30B 7/00 Single-crystal growth from solutions using solvents which are liquid at normal temperature, e.g. aqueous solutions from molten solvents C30B9/00; by normal or gradient freezing C30B11/00; under a protective fluid C30B27/00
• CPC Class Codes: G01N 2035/00336 Analysers adapted for operation in microgravity, i.e. spaceflight
• CPC Class Codes: G01N 25/14 by using distillation, extraction, sublimation, condensation, freezing, or crystallisation G01N25/02 takes precedence
• CPC Class Codes: Y10S 117/90 : Apparatus characterized by composition or treatment thereof, e.g. surface finish, surface coating
• CPC Class Codes: Y10T 117/10 : Apparatus
• CPC Class Codes: Y10T 117/1024 : for crystallization from liquid or supercritical state

Abstract

A device for promoting protein crystal growth (PCG) using microfluidic channels. A protein sample and a solvent solution are combined within a microfluidic channel having laminar flow characteristics which forms diffusion zones, providing for a well defined crystallization. Protein crystals can then be harvested from the device. The device is particularly suited for microgravity conditions.

Claims

1. A device for promoting protein crystallization growth from solution, comprising:

a body structure;

means located within said body structure for introduction of at least one solution containing protein and at least one solution containing a solvent; and

at least one microfluidic channel connected to said introduction means wherein said protein solution and said solvent solution interact to induce formation of protein crystals within said channel.

2. The device of claim 1, wherein said protein solution and said solvent solution flow laminarly in parallel contact within said microfluidic channel to establish a concentration gradient within said channel, allowing for protein crystallization.

3. The device of claim 1, wherein said protein solution introduction means and said solvent solution introduction means are each connected to said crystallization channel by a microfluidic channel.

4. The device of claim 3, wherein said protein microfluidic channel, said solvent microfluidic channel, and said crystallization channel form a T-Sensor structure.

5. The device of claim 1, further comprising a chamber coupled to said crystallization channel for harvesting said formed protein crystals.

6. The device of claim 1, further comprising fluid movement generating means coupled to said protein solution introduction means and said solvent solution introduction means for propelling said solutions through said crystallization channel.

7. The device of claim 6, wherein said fluid movement generating means comprises an air bellows.

8. The device of claim 1, further comprising a mixing means, coupled between said protein solution introduction means and said solvent introduction means interface and said crystallization channel for mixing said protein solution and said solvent solution completely to form a homogeneous mixture.

9. The device of claim 8, wherein said mixing means comprises a jet vortex mixer.

10. The device of claim 9, further comprising a solvent absorbing means coupled to said crystallization channel for absorbing solvent from said homogeneous mixture to increase the concentration of protein within said mixture, thereby inducing increased protein crystallization.

11. The device of claim 1, wherein said body structure is constructed from plastic.

12. A device for promoting protein crystallization growth from solution, comprising:

a body structure;

means located within said body structure for introduction of at least one solution containing protein, at least one solution containing a solvent, and at least one solution containing a combination of protein and a buffer;

a microfluidic structure coupled to said protein solvent introduction means and said combination protein and buffer solution introduction means for flowing said solutions laminarly in parallel to remove irreversible protein aggregates from said combined solutions;

and at least one microfluidic channel connected to said solvent introduction means and the output of said microfluidic structure wherein said solvent solution and said combined solutions interact to induce formation of protein crystals within said channel.

13. The device of claim 12, further comprising a waste chamber coupled to said microfluidic structure to retain said irreversible protein aggregates.

14. The device of claim 12, further comprising a chamber coupled to said crystallization chamber for harvesting said formed protein crystals.

15. The device of claim 12, further comprising fluid movement generating means coupled to said protein solution introduction means, said solvent solution introduction means, and said combined protein and buffer solution introduction means for propelling said solutions through said crystallization channel.

16. The device of claim 15, wherein said fluid movement generating means comprises an air pump.

17. The device of claim 12, wherein said microfluidic structure comprises an H-filter.