Claims
- 1. A method of stabilizing an aqueous enzyme comprising the step of contacting the enzyme selected from the group consisting of peroxidase enzymes, alkaline phosphatase enzymes and antibody conjugates thereof with an aqueous solution of a protein stabilizer additive comprising:
- a. a tris(hydroxymethyl)methyl compound of formula 1;
- (HOCH.sub.2).sub.3 C--R (1)
- wherein R is: NH.sub.2 ; NR.sup.1 R.sup.2 wherein R.sup.1 and R.sup.2 may be independently: H, C.sub.1 -C.sub.4 alkyl, C.sub.1 -C.sub.4 hydroxyalkyl, C.sub.1 -C.sub.4 alkyl sulfonate, C.sub.1 -C.sub.4 hydroxyalkyl sulphonate; C.sub.1 -C.sub.4 alkyl NHC(CH.sub.2 OH).sub.3 ; C.sub.1 -C.sub.4 alkyl carboxylate;
- b. a cationic polyelectrolyte selected from the group consisting of copolymers of vinyl pyrollidone and quaternary methyl methacrylate and DEAE dextran;
- c. a polyol selected from the group consisting of glycerol, ethylene glycol, sorbitol and lactitol; and
- d. one or more salts selected from the group consisting of salt of calcium, magnesium and cobalt.
- 2. A method as claimed in claim 1 wherein the tris(hydroxymethyl)) methyl compound is selected from the group consisting of: tris(hydroxymethyl)aminomethane and salts thereof selected from the group consisting of chloride, maleate, phosphate, succinate salts; 1,3-bis{tris(hydroxymethyl)methylamino}propane; bis(2-hydroxyethyl)amino-tris(hydroxymethyl)methane; N-{tris(hydroxymethyl)methyl}-2-aminoethane sulfonate; N-{tris(hydroxymethyl)methyl}-3-aminopropane sulfonate; N-{tris(hydroxymethyl)methyl}-3-amino-2-hydroxypropane sulfonate; and N-{tris(hydroxymethyl)methyl}-glycine.
- 3. A method as claimed in claim 1 wherein the cationic polyelectrolyte has a molecular weight of 150 to 5,000,000 Daltons.
- 4. A method as claimed in claim 1 wherein the cationic polyelectrolyte is present in an amount of 0.01 to 10% w/v.
- 5. A method as claimed in claim 1 wherein the polyol is present in an amount of 0.1 to 25% w/v.
- 6. A method as claimed in claim 1 wherein said peroxidase enzyme is selected from the group consisting of horseradish peroxidase and arthromyces peroxidase.
- 7. A method as claimed in claim 2, wherein the tris(hydroxymethyl) compound is selected from the group consisting of tris(hydroxymethyl)aminomethane and salts thereof selected from the group consisting of chloride, maleates, phosphate, succinate salts; 1,3-bis{tris(hydroxymethyl)methylamino}propane; and bis(2-hydroxyethyl)aminotris(hydroxymethyl)) methane.
- 8. A method as claimed in claim 1, wherein said enzyme is horseradish peroxidase, said tris is 20 mM tris/HCL buffer pH 8.0, said polyol is ethylene glycol, said cationic polyelectrolyte is DEAE-dextran (MW 500K), and said salt is calcium chloride.
- 9. A method as claimed in claim 8, wherein at least 99% of the enzyme's activity is retained after 33 days at 37.degree. C.
Priority Claims (1)
Number |
Date |
Country |
Kind |
9320782 |
Oct 1993 |
GBX |
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Parent Case Info
This application is a 371 of PCT/GB94/02180 filed Oct. 6, 1994.
PCT Information
Filing Document |
Filing Date |
Country |
Kind |
102e Date |
371c Date |
PCT/GB94/02180 |
10/6/1994 |
|
|
7/1/1996 |
7/1/1996 |
Publishing Document |
Publishing Date |
Country |
Kind |
WO95/10605 |
4/20/1995 |
|
|
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