Gel plates, equipment and kits for combined electrophoretic-immunoelectrophoretic analysis

• US 5,238,651 A
• Filed: 07/23/1990
• Issued: 08/24/1993
• Est. Priority Date: 07/23/1990
• Status: Expired due to Fees

• Current Assignee: New York University New York NY
• Original Assignee: New York University
• Inventors: Chuba, Joseph V.
• Primary Examiner(s): Housel, James C.
• Assistant Examiner(s): Snay, Jeffrey R.
• Application Number:
  US07/555,874
• Time in Patent Office: 1,128 Days
• Field of Search: 436/514, 436/515, 436/516, 436/149, 204/182.8, 204/182.7, 204/182.9, 204/299 R, 204/301, 204/183.3, 204/180.1, 83/679, 83/651, 83/648, 422/61, 422/101, 422/104
• US Class Current: 422/430
• CPC Class Codes: B01D 57/02 by electrophoresis treatment of water, waste water, sewage or sludge by electrophoresis C02F1/469; electrophoretic production of compounds or non-metals C25B7/00; investigating or analysing materials by using electrophoresis G01N27/26
• CPC Class Codes: G01N 27/44717 Arrangements for investigating the separated zones, e.g. localising zones
• CPC Class Codes: G01N 27/44721 by optical means
• CPC Class Codes: G01N 27/44739 Collecting the separated zones, e.g. blotting to a membrane or punching of gel spots
• CPC Class Codes: G01N 27/44743 Introducing samples

Abstract

Combined electrophoretic-immunoelectrophoretic analysis is conducted using a gel plate having a plurality of preformed trenches therein dividing the plate into a number of electrophoretic tracks. Serum samples are applied to the tracks such that each pair of tracks has the same sample. After subjecting the tracks to electrophoresis, one track from each pair of matching tracks is removed for protein staining by inserting a spatula into a trench and under the track to be removed. Small dabs of high vacuum silicone stopcock grease are applied to the ends of each sidewall of the remaining gel tracks and antiserum is applied therebetween. After immunoprecipitation and staining, the removed tracks are returned to their original positions to permit exact correlation of said immunoprecipitation and electrophoresis patterns. This analysis technique may be facilitated by use of equipment which includes a plate holder, a plate holder frame, underlay inserts, left and right margin inserts, a guide bar frame, templates, spatulas and plate handling hooks, burnishing rods and applicator pens. The trenches may be formed in conventional gel plates by specially designed equipment including a trench former insert frame, a double bladed gel cutter and a gel displacement beak.

Claims

1. A kit for facilitating combined electrophoretic-immunoelectrophoretic analysis, comprising:

1) a plate holder comprising side walls defining a predetermined perimeter, gel-support surface means for supporting a gel plate, and guide means for guiding a gel plate into a predetermined position on said gel-support surface means; and

2) a plate holder frame comprising guide wall means having an inner perimeter corresponding to the outer perimeter of said plate holder side walls for holding said plate holder therewithin, plate holder support surface means for supporting said plate holder within said wall means, and a peripheral surface disposed outside of said guide wall means, said peripheral surface being an extension of said plate holder support surface means, said peripheral surface extending beyond said guide wall means for a distance sufficient to serve as a hand support when in use.

2. A kit in accordance with claim 1, and further including left and right gel plate holding support means for resting on said peripheral surface of said plate holder frame and simultaneously holding a gel plate in place on said plate holder when in use, each of said gel plate holding support means comprising a hand support surface sized and dimensioned so as to rest upon a portion of said peripheral surface of said plate holder frame, a plate contact means for resting upon a gel plate when a gel plate is in place in said plate holder frame, and bridge means connecting said hand support means and said gel holding means for bridging said side walls of said plate holder and said guide wall means of said plate holder frame.

3. A kit in accordance with claim 1, and further including a guide bar frame comprising side wall means having an outer perimeter sized and dimensioned so as to precisely fit within the inner perimeter of the side walls of said plate holder, and guide bar means traversing said guide bar frames so as to be suspended above a gel plate when in place on said gel holder when said guide bar frame is disposed within said plate holder, said guide bar means being precisely disposed in a predetermined position so as to facilitate a step of the electrophoretic-immunoelectrophoretic analysis method to be performed by means of the kit.

4. A kit in accordance with claim 1, and further including spatula means for transferring tracks of gel matrix to and from a gel plate when the kit is in use.

5. A kit in accordance with claim 1, and further including

a trench former insert frame comprising perimeter walls having an outer perimeter sized and dimensioned so as to fit precisely within the inner perimeter of the side walls of said plate holder, and two matching rows of multiple slots disposed on opposing portions of said perimeter walls, each of said pairs of slots being precisely disposed in a predetermined position with respect to a gel plate held in said plate holder when said trench former insert frame is disposed in said plate holder;

a double bladed trench cutter comprising two gel cutting blades, two blade support means disposed parallel to and in predetermined distance from one another, each having one of said cutting blades mounted thereon on the inside surface facing the other blade support means, the blade portion of said cutting blades extending out beneath said blade support means, for supporting said blades in a predetermined position, and two thumb plate means connected to said blade support means on the side thereof opposite the side from which said blades extend and fixing said blade support means and the supported blades such that said blades are held parallel to one another with a fixed predetermined separation therebetween and such that the space between said blades between said thumb plate means remains completely accessible from the side of said blade support means opposite the side from which said blades extend, and wherein the outside dimensions of said blade support means precisely correspond to the dimensions of said multiple slots so as to precisely fit therewithin when in use; and

a gel displacement beak having a width exactly equal to the space between said blades, said gel displacement beak having a beak portion tapering to a tip at one end and having a handle portion at the other end.

6. A kit in accordance with claim 1, and further including:

a trench former frame shaped and dimensioned so as to cooperate with said plate holder and be maintained in a predetermined position with respect to a gel plate supported by said plate holder when in use;

a double bladed trench cutter comprising

(a) two gel cutting blades

(b) two blade support means disposed parallel to and at a predetermined distance from one another, each having one of said cutting blades mounted thereon on the inside surface facing the other blade supporting means, the blade portion of said cutting blades extending out beneath said blade support means, for supporting said blades in a predetermined position, and

(c) connecting means connected to said blade support means and fixing said blade support means and the supported blades such that the space between said blades remains accessible from the side of said blade support means opposite the side from which said blades extend; and

positioning means associated with said trench former frame and said trench cutter for ensuring that said trench cutter is positioned on said trench former frame at a predetermined position with respect to the gel plate to be trenched when in use.

7. A kit in accordance with claim 1, wherein said plate holder support surface means has a recess therein sized and dimensioned so as to receive and hold in position an underlay insert having printed matter which is desired to be seen through said plate holder and any gel plate disposed thereon when in use, and wherein said gel support surface means of said plate holder is substantially transparent.

8. A kit in accordance with claim 7, and further including a plurality of underlay inserts sized and dimensioned so as to fit and be held in said recess of said plate holder support surface means, each of said inserts showing or explaining one or more steps of the combined electrophoretic-immunoelectrophoretic analysis process which is to be performed using the kit.

9. A kit in accordance with claim 1, wherein said gel support surface means of said plate holder includes an opening therein disposed so as to be traversed by an edge of a gel plate when in position in said plate holder.

10. A kit in accordance with claim 9, and further including a plate handling hook including a handle at one end and hook means at the other end for insertion into the opening in the gel support surface means of said plate holder such that the hook means can reach beneath a gel plate in place on said plate holder when in use and facilitate the lifting of the gel plate from said plate holder.

11. A kit in accordance with claim 1, and further including a plurality of gel plates each comprising:

a substantially rigid, electrically non-conducting, non-porous backing; and

a layer of a substantially non-seiving neutral electrophoresis matrix adhered to said backing, said matrix having a plurality of parallel trenches therein extending linearly from points at or near the edges of the gel, said trenches having a width of about 0.5 to about 3 mm and being spaced from one another at a distance approximately equal to the width of a desired electrophoresis track, thereby dividing the gel into a plurality of tracks, no other gaps in said matrix being present, the side walls of said trenches being perpendicular to said backing and extending vertically all the way to said backing.

12. A kit in accordance with claim 11, and further including a template having a number of colinear slits, said slits being arranged such that when said template is disposed on said gel plate, each said slit is perpendicular to the longitudinal direction of said trenches and disposed in a predetermined position between two adjacent trenches.

13. A kit for facilitating the formation of precisely disposed trenches in a gel plate, comprising:

a trench former insert frame comprising perimeter walls and two matching rows of multiple slots disposed on opposing portions of said perimeter walls each of said pairs of slots being precisely disposed in a predetermined position;

a double bladed trench cutter comprising two gel cutting blades, two blade support means disposed parallel to and in predetermined distance from one another, each having one of said cutting blades mounted thereon on the inside surface facing the other blade support means, the blade portion of said cutting blades extending out beneath said blade support means, for supporting said blades in a predetermined position, and two thumb plate means connected to said blade support means on the side thereof opposite the side from which said blades extend and fixing said blade support means and the supported blades such that said blades are held parallel to one another with a fixed predetermined separation therebetween and such that the space between said blades between said thumb plate means remains completely accessible from the side of said blade support means opposite the side from which said blades extend, and wherein the outside dimensions of said blade support means precisely correspond to the dimensions of said multiple slots so as to precisely fit therewithin when in use; and

a gel displacement beak having a width exactly equal to the space between said blades, said gel displacement beak having a beak portion tapering to a tip at one end and having a handle portion at the other end.

14. A kit in accordance with claim 13, and further including a plate holder comprising side walls defining a predetermined perimeter, the inner circumference of which exactly corresponds to the outer perimeter of the perimeter walls of said trench former insert frame, gel support surface means for supporting a gel plate, and guide means for guiding a gel plate into a predetermined position on said gel support surface means.

15. A kit for use in preparing a trenched gel plate, comprising:

a trench former frame which is disposed in a predetermined position with respect to the gel plate to be trenched when in use;

a double bladed trench cutter comprising

a) two gel cutting blades,

b) two blade support means disposed parallel to and at a predetermined distance from one another, each having one of said cutting blades mounted thereon on the inside surface facing the other blade supporting means, the blade portion of said cutting blades extending out beneath said blade support means, for supporting said blades in a predetermined position, and

c) connecting means connected to said blade support means and fixing said blade support means and the supported blades such that the space between said blades remains accessible from the side of said blade support means opposite the side from which said blades extend; and

positioning means associated with said trench former frame and said trench cutter for ensuring that said trench cutter is positioned on said trench former frame at a predetermined position with respect to the gel plate to be trenched when in use.

16. A kit in accordance with claim 15, wherein said positioning means comprises two matching rows of multiple slots disposed on opposing portions of said trench former frame, each of said pairs of slots being precisely disposed in a predetermined position with respect to the gel plate to be trenched when in use, and wherein the outside dimensions of said blade support means of said trench cutter precisely corresponds to the dimensions of said multiple slots so as to precisely fit therewithin when in use.

17. A kit in accordance with claim 15, wherein said connecting means comprise two thumb plate means connected to said blade support means on the side thereof opposite the side from which said blades extend and fixing said blade support means and the supported blades such that said blades are held parallel to one another with a fixed predetermined separation therebetween and such that the space between said blades between said thumb plate means remains completely accessible from the side of said blade support means opposite the side from which said blades extend.