Chromatographic column

• US 4,740,306 A
• Filed: 03/17/1986
• Issued: 04/26/1988
• Est. Priority Date: 03/17/1986
• Status: Expired due to Fees

• Current Assignee: Temple University of The Commonwealth System of Higher Education A Nonprofit Organization of PA
• Original Assignee: Temple University of The Commonwealth System of Higher Education
• Inventors: Kirsch, Ted M., Litwack, Gerald
• Primary Examiner(s): Therkorn, Ernest G.
• Application Number:
  US06/840,399
• Time in Patent Office: 771 Days
• Field of Search: 210/635, 210/656, 210/657, 210/659, 210/198.2, 210/283, 210/290, 55/67, 55/386, 436/501, 435/219, 435/317, 435/320, 435/803, 435/815, 530/395, 530/396, 530/413, 530/417
• US Class Current: 210/198.2
• CPC Class Codes: B01D 15/36 involving ionic interaction
• CPC Class Codes: B01D 15/362 Cation-exchange
• CPC Class Codes: B01J 39/26 Cation exchangers for chromatographic processes
• CPC Class Codes: G01N 30/02 Column chromatography
• CPC Class Codes: G01N 30/52 Physical parameters
• CPC Class Codes: G01N 30/60 Construction of the column
• CPC Class Codes: Y10S 435/803 Physical recovery methods, e.g. chromatography, grinding
• CPC Class Codes: Y10S 435/815 by sorption

Abstract

A chromatographic column and process for isolating or purifying steroid hormone or cell membrane receptors using the column are provided in which the column contains at least three resin layers between the inlet and the outlet ends of the column, the layer closest the inlet being a strong cationic exchange resin, the middle layer being a matrix containing a triazine dye that will bind proteases and proteins with dinucleotide fold conformations, and the layer closest the outlet being a weak anionic exchange resin, the ratio of the volumes of the resin layers being 1:2:1.

Claims

1. A chromatographic column having at least three resin layers between the inlet and the outlet ends of the column, the layer closest the inlet being a strong cationic exchange resin, the middle layer being a backbone containing a triazine dye that will bind proteases and proteins with dinucleotide fold conformations, and the layer closest to the outlet being a weak anionic exchange resin, the ratio of the volumes of the resin layers being 1:

2;

1.

2. The chromatographic column of claim 1 wherein the resin layer closest to the inlet is a six percent phosphorylated agarose gel crosslinked with epichlorohydrin.

3. The chromatographic column of claim 1 wherein the middle resin layer is a monochlorotriazine dye-containing synthetic matrix.

4. The chromatographic column of claim 3 wherein the matrix is a free radical polymerization product of N-acryloyl-2-amino-2-hydroxymethyl-1,3-propanediol.

5. The chromatographic column of claim 1 wherein the resin layer closest to the outlet is a diethylaminoethyl-containing synthetic matrix.

6. The chromatographic column of claim 4 wherein the matrix is a free radical polymerization product of N-acryloyl-2-amino-2-hydroxymethyl-1,3-propanediol.

7. The chromatographic column of claim 1 wherein the ratio of the volumes of the resin layers is 20:

40;

20.

8. A construction for rapid one-step isolation and purification of activated and unactivated forms of steroid hormone and cell membrane receptors which comprises a chromatographic column containing a matrix of resin layers between the inlet and the outlet, the resin layer closest to the inlet being a strong cationic exchange resin, the middle layer being a triazine dye containing exchange resin and the layer closest to the outlet being a weak anionic exchange resin, the ratio of the volumes of the resin layers being 1:

2;

1.

9. The construction of claim 8 wherein the resin layer closest to the inlet is a six percent phosphorylated agarose gel crosslinked with epichlorohydrin.

10. The construction of claim 8 wherein the middle resin layer is a monochlorotriazine dye-containing synthetic matrix.

11. The construction of claim 10 wherein the matrix is a free radical polymerization product of N-acryloyl-2-amino-2-hydroxymethyl-1,3-propanediol.

12. The construction of claim 8 wherein the resin layer closest to the outlet is a diethylaminoethyl-containing synthetic matrix.

13. The construction of claim 12 wherein the matrix is a free radical polymerization product of N-acryloyl-2-amino-2-hydroxymethyl-1,3-propanediol.