Gel electroelution and sample separation devices and associated processes

Abstract

A gel electroelution device can have a gel spot column having upstream and downstream openings in fluid communication with an inlet and outlet, respectively. The gel spot column receives a gel spot and negative and positive electrodes are fluid communication with the inlet and outlet, respectively. A gel electroelution process can involve flowing a buffer solution through the gel spot in a first direction, and creating an electric field across the gel spot in the same direction. A separator device can have a generally cylindrical collection reservoir in fluid communication with inlet, filtrate, and retentate ports, a filter intermediate the inlet and filtrate ports. The inlet and retentate ports can intersect the collection reservoir in a manner to induce a cyclonic flow. A flow separation process can involve inducing a generally cyclonic flow in the collection reservoir, and filtering the fluid therein to retain the retentate.

Description

BRIEF DESCRIPTION OF THE DRAWING FIGURES

A more complete understanding of the gel electroelution and sample separation processes and devices can be obtained by considering the following detailed description in conjunction with the accompanying drawing figures, in which:

FIG. 1 is an exploded, front perspective view of an embodiment of a gel electroelution device;

FIG. 2 is an exploded, rear perspective view of the gel electroelution device shown in FIG. 1;

FIG. 3 is a side sectional view of the gel electroelution device shown in FIG. 1;

FIG. 4 is a side sectional view of another embodiment of a gel electroelution device;

FIG. 5 is a perspective view of an embodiment of a separator device;

FIG. 6 is a horizontal sectional view of the separator device shown in FIG. 5.

FIG. 7 is a vertical sectional view of the separator device shown in FIG. 5.

FIG. 8 is front perspective view of an embodiment of a combined gel electroelution and separator device.

FIG. 9 is a rear perspective view of the combined gel electroelution and separator device shown in FIG. 8.

FIG. 10 is a bottom perspective view of the combined gel electroelution and separator device shown in FIG. 8, with the bottom housing removed.

Claims

1. A gel electroelution process to extract electrophoretically separated intact proteins from a gel matrix, said process comprising: a. immobilizing a gel spot from said gel matrix, said gel spot containing said proteins to be extracted;b. flowing a buffer solution through said gel spot in a first direction; andc. creating an electric field across said gel spot in said first direction, such that said proteins electrophoretically migrate out of said gel spot and into said buffer solution.

2. The gel electroelution process of claim 1 further comprising preventing admixing of flow on upstream and downstream sides of said gel spot.

3. The gel electroelution process of claim 1 wherein said gel matrix is an SDS-PAGE gel matrix, and said buffer solution mimics a natural running buffer of said SDS-PAGE gel separation.

4. The gel electroelution process of claim 3 wherein said SDS-PAGE gel matrix has not been chemically fixed prior to staining.

5. The gel electroelution process of claim 1 further comprising filtering said buffer solution downstream of said gel spot to remove particles of gel.

6. The gel electroelution process of claim 1 further comprising collecting migrated proteins in a reservoir.

7. The gel electroelution process of claim 1 further comprising collecting the migrated proteins in a solid phase extraction trapping column.

8. A gel electroelution device comprising: a. a gel spot column having upstream and downstream openings, said gel spot column adapted to receive therein a gel spot containing proteins to be extracted therefrom;b. an inlet in fluid communication with said upstream opening of said gel spot column;c. an outlet in fluid communication with said downstream opening of said gel spot column;d. a negative electrode in fluid communication with said inlet;e. a positive electrode in fluid communication with said outlet; andf. wherein said positive and negative electrodes are operable to create an electric field across said gel spot in the same direction as a flow of buffer solution from said inlet through said outlet in order to electrophoretically migrate said proteins out of said gel spot and into said buffer solution.

9. The gel electroelution device of claim 8 further comprising a frit plug intermediate said gel spot and said outlet.

10. The gel electroelution device of claim 8 further comprising a purge line in fluid communication with said gel spot column intermediate said gel well and said outlet.

11. The gel electroelution device of claim 8 further comprising a housing, and said gel spot column removably enclosed in said housing.

12. The gel electroelution device of claim 11 wherein said gel spot column further comprises a gel spot cutter, such that when said gel spot cutter is used to cut a gel spot, said gel spot is retained in said gel spot cutter, and said gel spot cutter is then enclosable in said housing with said gel spot retained therein.

13. A separation process comprising: a. flowing a fluid sample into collection reservoir;b. inducing a generally cyclonic flow in said collection reservoir;c. filtering said fluid sample using hydrodynamic pressure created in said collection reservoir; andd. retaining retentate in said collection reservoir.

14. The separation process of claim 13 further comprising: a. flowing a solution into said collection reservoir containing said retentate; andb. flowing said solution and retentate out of said collection reservoir.

15. The separation process of claim 1 further comprises at least one of rinsing, desalting, purifying, and concentrating said retentate in said collection reservoir.

16. The separation process of claim 1 further comprising fluorescent labeling of said retentate in said collection reservoir.

17. A separator device comprising: a. a collection reservoir in fluid communication with an inlet port, a filtrate port, and a retentate port; andb. a filter receptacle intermediate said inlet port and said filtrate port adapted to receive therein a filter.

18. The separator device of claim 17 wherein said collection reservoir is generally cylindrical, and each of said inlet port and retentate port intersect said collection reservoir in an off-center generally orthogonal orientation such that a cyclonic flow is induced in said collection reservoir.

19. The separator device of claim 17 wherein said inlet, retentate and filtrate ports are marked with different colors.