Glyceryl agarose and borate compositions

• US 5,212,299 A
• Filed: 05/10/1991
• Issued: 05/18/1993
• Est. Priority Date: 05/10/1991
• Status: Expired due to Term

• Current Assignee: Cambrex Bio Science Rockland, Inc. (Lonza Group AG)
• Original Assignee: FMC Corporation
• Inventors: Smith, Joshua D.
• Primary Examiner(s): GRIFFIN, RONALD
• Application Number:
  US07/698,228
• Time in Patent Office: 739 Days
• Field of Search: 106/162, 106/205, 536/3, 536/115, 536/114, 514/54, 530/120, 436/529
• US Class Current: 536/114
• CPC Class Codes: C08B 37/0039 Agar; Agarose, i.e. D-galactose, 3,6-anhydro-D-galactose, methylated, sulfated, e.g. from the red algae Gelidium and Gracilaria; Agaropectin; Derivatives thereof, e.g. Sepharose, i.e. crosslinked agarose
• CPC Class Codes: C08K 3/38 Boron-containing compounds
• CPC Class Codes: C08L 5/12 Agar or agar-agar , i.e. mixture of agarose and agaropectin; Derivatives thereof
• CPC Class Codes: G01N 27/44747 Composition of gel or of carrier mixture

Abstract

Aqueous gel compositions and their use in electrophoresis, and anhydrous compositions useful in the preparation of the aqueous gel compositions.

The aqueous gel compositions comprise:

A. water,

B. at least one di- or tri-hydroxy C_2-4 alkyl ether moiety (preferably glyceryl) substituted agarose, and

C. from about 20 to about 400 mM of at least one borate compound per liter of aqueous gel composition.

The anhydrous compositions comprise the agarose in powder form and the borate in powder or aqueous solution.

Claims

1. An aqueous gel composition characterized in that it comprises:

(A) at least one di-or tri-hydroxy alkyl ether moiety substituted gel-forming agarose present in a gel-forming effective amount;

(B) at least one borate compound other than a strongly reducing or strongly axoidizing compound present in a gel-handling-effective amount; and

(C) water sufficient to form a gel.

2. The composition of claim 1 characterized in that the moiety is glyceryl.

3. The composition of claim 2 characterized in that said at least one borate compound is present in about 20 to 400 mM of aqueous gel composition.

4. The composition of claim 3 characterized in that said at least one borate compound is present in about 40 to 150 mM of aqueous gel composition.

5. The composition of claim 4 characterized in that said at least one borate compound is present in about 50 to 100 mM of aqueous gel composition.

6. The composition of claim 2 characterized in that it has a pH of about 3.0 to 12.0.

7. The composition of claim 6 characterized in that it has a pH of about 6.0 to 10.0.

8. The composition of claim 7 characterized in that it has a pH of about 7.5 to 9.5.

9. The composition of claim 8 characterized in that it has a pH of about 8.0 to 8.5.

10. The composition of claim 2 characterized in that said at least one glyceryl agarose is present in about 0.1 to 15% weight/volume.

11. The composition of claim 10 characterized in that said at least one glyceryl agarose is present in about 0.5 to 8% weight/volume.

12. The composition of claim 11 characterized in that said at least one glyceryl agarose is present in about 1 to 6% weight/volume.

13. The composition of claim 2 characterized in that the borate compound is at least one of BH₃ O₃, alkali metal borate salts, alkaline earth metal borate salts, alkali metal organoboron compounds, and alkaline earth metal organoboron compounds.

14. The composition of claim 13 characterized in that the borate compound is BH₃ O₃.

15. The composition of claim 2 characterized in that the glyceryl agarose contains about 0.08 to 1.3 glyceryl moieties per each agarose agarobiose unit.

16. The composition of claim 15 characterized in that the glyceryl agarose contains about 0.10 to 0.75 glyceryl moieties per each agarose agarobiose unit.

17. The composition of claim 2 characterized in that the glyceryl agarose is based on agarose derived from R. Gelidium or R. Pterocladia.

18. The composition of claim 2 characterized in that the pH of the composition is about 6.0 to about 10.0, and the at least one borate compound is BH₃ O₃ present in about 50 to about 150 mM per liter of composition.

19. The composition of claim 2 characterized in that the pH is about 7.5 to 9.5, the at least one borate compound is BH₃ O₃ present in about 80 to about 100 mM of composition, and the glyceryl agarose is present in about 1 to about 6% weight/volume.

20. A kit for the preparation of the composition of claim 2 upon admixture with water characterized in that it comprises measured amounts of:

(A) said at least one glyceryl agarose; and

(B) said at least one borate compound.

21. The kit of claim 20 characterized in that the borate compound is BH₃ O₃.

22. The kit of claim 21 characterized in that the glyceryl agarose has about 0.08 to 1.3 glyceryl moieties per each agarose agarobiose unit.

23. A method for gel electrophoresis characterized in that the gel used therein is an aqueous gel composition comprising:

(A) at least one di- or tri-hydroxy C_2-4 alkyl ether moiety substituted agarose present in a gel-forming-effective amount;

(B) at least one borate compound present in a gel-handling-effective amount; and

(C) water sufficient to form a gel.

24. The method of claim 23 characterized in that the moiety is glyceryl.

25. The method of claim 24 characterized in that the pH of the gel composition is about 6.0 to 10.0, and the borate compound is BH₃ O₃ present in about 50 to about 150 mM per liter of gel composition.

26. A method for imparting gel-handling properties to an aqueous glyceryl agarose gel characterized by admixing therewith a handling-effective amount of at least one borate prior to, during, or subsequent to, the formation of said gel.

27. The method of claim 26 characterized in that the pH of the gel composition is about 6.0 to 10.0, and the borate compound is BH₃ O₃ present in about 50 to about 150 mM per liter of gel composition.