Microfluidic protein crystallography

• US 8,691,010 B2
• Filed: 04/15/2011
• Issued: 04/08/2014
• Est. Priority Date: 06/28/1999
• Status: Expired due to Fees

• Current Assignee: California Institute of Technology
• Original Assignee: California Institute of Technology
• Inventors: Hansen, Carl L., Sommer, Morten, Quake, Stephen R.
• Primary Examiner(s): Kunemund, Bob M
• Application Number:
  US13/087,697
• Publication Number:
  US 20110306522A1
• Time in Patent Office: 1,089 Days
• Field of Search: 117/68, 117/69, 117/70
• US Class Current: 117/68
• CPC Class Codes: B01J 19/0046 Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays synthesis methods per se C40B50/00
• CPC Class Codes: B01J 2219/00286 Reactor vessels with top and bottom openings
• CPC Class Codes: B01J 2219/00355 peristaltic
• CPC Class Codes: B01J 2219/00378 Piezoelectric or ink jet dispensers
• CPC Class Codes: B01J 2219/00396 Membrane valves
• CPC Class Codes: B01J 2219/00398 in multiple arrangements
• CPC Class Codes: B01J 2219/00439 using micromirror arrays
• CPC Class Codes: B01J 2219/005 Beads
• CPC Class Codes: B01J 2219/00527 Sheets
• CPC Class Codes: B01J 2219/0059 Sequential processes
• CPC Class Codes: B01J 2219/00605 the compounds being directly bound or immobilised to solid supports
• CPC Class Codes: B01J 2219/0061 The surface being organic
• CPC Class Codes: B01J 2219/00612 the surface being inorganic
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00704 integrated with the reactor apparatus
• CPC Class Codes: B01J 2219/00707 separated from the reactor apparatus
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01J 2219/00725 Peptides
• CPC Class Codes: B01J 2219/00756 Compositions, e.g. coatings, crystals, formulations
• CPC Class Codes: B01L 2200/025 Align devices or objects to ensure defined positions relative to each other
• CPC Class Codes: B01L 2200/027 : for microfluidic devices
• CPC Class Codes: B01L 2200/0605 : Metering of fluids
• CPC Class Codes: B01L 2200/0673 : Handling of plugs of fluid surrounded by immiscible fluid
• CPC Class Codes: B01L 2200/10 : Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
• CPC Class Codes: B01L 2300/0681 : Filter
• CPC Class Codes: B01L 2300/0861 : Configuration of multiple channels and/or chambers in a single devices
• CPC Class Codes: B01L 2300/123 : Flexible; Elastomeric
• CPC Class Codes: B01L 2300/14 : Means for pressure control
• CPC Class Codes: B01L 2300/18 : Means for temperature control
• CPC Class Codes: B01L 2400/0472 : Diffusion
• CPC Class Codes: B01L 2400/0481 : squeezing of channels or chambers
• CPC Class Codes: B01L 2400/0655 : pinch valves
• CPC Class Codes: B01L 2400/0688 : surface tension valves, capillary stop, capillary break
• CPC Class Codes: B01L 3/06 : Crystallising dishes
• CPC Class Codes: B01L 3/5027 : by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• 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 3/502738 : characterised by integrated valves microvalves F16K99/0001
• CPC Class Codes: B01L 7/54 : using spatial temperature gradients
• CPC Class Codes: B01L 9/527 : for microfluidic devices, e.g. used for lab-on-a-chip
• CPC Class Codes: C30B 29/58 : Macromolecular compounds
• CPC Class Codes: C30B 35/00 : Apparatus not otherwise provided for, specially adapted for the growth, production or after-treatment of single crystals or of a homogeneous polycrystalline material with defined structure
• 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: C30B 7/10 : by application of pressure, e.g. hydrothermal processes
• CPC Class Codes: C30B 7/14 : the crystallising materials being formed by chemical reactions in the solution
• CPC Class Codes: C40B 40/10 : Libraries containing peptides or polypeptides, or derivatives thereof
• CPC Class Codes: F04B 43/043 : Micropumps
• CPC Class Codes: F04B 43/14 : having plate-like flexible members
• CPC Class Codes: F16K 2099/0074 : using photolithography, e.g. etching
• CPC Class Codes: F16K 2099/0078 : using moulding or stamping
• CPC Class Codes: F16K 2099/008 : Multi-layer fabrications
• CPC Class Codes: F16K 2099/0094 : Micropumps
• CPC Class Codes: F16K 99/0001 : Microvalves microdevices B81B1/00; manufacture or treatment of devices or systems in or on a substrate B81C1/00; microfluidic structures B01L3/5027; micropumps F04B19/006
• CPC Class Codes: F16K 99/0015 : Diaphragm or membrane valves
• CPC Class Codes: F16K 99/0028 : Valves having multiple inlets or outlets
• CPC Class Codes: F16K 99/0057 : the fluid being the circulating fluid itself, e.g. check valves
• CPC Class Codes: G01N 2013/003 : Diffusion; diffusivity between liquids
• CPC Class Codes: Y10T 117/10 : Apparatus
• CPC Class Codes: Y10T 117/1004 : with means for measuring, testing, or sensing
• CPC Class Codes: Y10T 117/1008 : with responsive control means
• CPC Class Codes: Y10T 117/1024 : for crystallization from liquid or supercritical state

Abstract

The use of microfluidic structures enables high throughput screening of protein crystallization. In one embodiment, an integrated combinatoric mixing chip allows for precise metering of reagents to rapidly create a large number of potential crystallization conditions, with possible crystal formations observed on chip. In an alternative embodiment, the microfluidic structures may be utilized to explore phase space conditions of a particular protein crystallizing agent combination, thereby identifying promising conditions and allowing for subsequent focused attempts to obtain crystal growth.

Claims

1. An apparatus for investigating crystallization comprising:

a microfluidic formulator comprising a microfluidic chamber configured to receive a crystallization target and a precipitant;

a light source configured to illuminate the microfluidic chamber; and

a light detector configured to receive light transmitted through the microfluidic chamber.

2. The apparatus of claim 1 wherein the microfluidic chamber comprises a rotary flow chamber.

3. The apparatus of claim 1 further comprising a peristaltic pump actuable to flow the crystallization target and crystallizing agent through the rotary flow chamber.

4. The apparatus of claim 1 wherein the detector comprises a plurality of pixels configured to receive light transmitted through a portion of the rotary flow chamber.

5. The apparatus of claim 1 wherein the detector comprises a charge coupled device.

6. The apparatus of claim 2 further comprising a processor configured to receive an electronic signal from the light detector indicating an intensity of light transmitted through the chamber and received on the pixel, the processor comprising a computer readable storage medium having recorded instructions to calculate from the electronic signal a standard deviation of pixel intensity of light transmitted through the portion, and to compare the standard deviation to a background value recorded in an absence of the crystallization target.

10. The apparatus of claim 3 wherein the peristaltic pump comprises a plurality of peristaltic pump valves.

11. The apparatus of claim 10 wherein the plurality of peristaltic pump valves comprises are positioned in series along a channel that is in fluidic communication with the rotary flow chamber.

7. An apparatus for investigating crystallization comprising:

a chip holder for holding a microfluidic chamber configured to receive a crystallization target and a precipitant;

a light source configured to illuminate the microfluidic chamber; and

a light detector configured to receive light transmitted through the microfluidic chamber.

8. The apparatus of claim 7 wherein the detector comprises a charge coupled device.

9. The apparatus of claim 7 further comprising a processor adapted to receive an electronic signal from the light detector indicating an intensity of light transmitted through a microfluidic chamber.