Refolding transforming growth factor beta family proteins
First Claim
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1. A method of inducing folding of a denatured polypeptide, the method comprising:
- providing a denatured polypeptide comprising a transforming growth factor beta (TGF beta) superfamily member; and
contacting the polypeptide with an amount of a refolding buffer effective to induce folding of the polypeptide, wherein the refolding buffer comprises (i) potassium phosphate or sodium phosphate at a concentration of 25 mM to 150 mM with a pH of 5.8 to 8.0, (ii) guanidine-HCl at a concentration of 0.3 M to 2 M, (iii) L-Arginine at a concentration of 0.25 M to 1 M, (iv) Tween-80 at a concentration of 0.05% to 1%, and (v) oxidized glutathione at a concentration of 1 mM to 4 mM and reduced glutathione at a concentration of 0.05 mM to 0.8 mM, wherein the ratio of oxidized to reduced glutathione is from 5;
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Abstract
Compositions and methods for folding proteins belonging to the transforming growth factor beta superfamily are disclosed. The compositions and methods allow for the folding of such proteins when produced in an expression system that does not yield a properly folded, biologically active product.
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Citations
36 Claims
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1. A method of inducing folding of a denatured polypeptide, the method comprising:
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providing a denatured polypeptide comprising a transforming growth factor beta (TGF beta) superfamily member; and contacting the polypeptide with an amount of a refolding buffer effective to induce folding of the polypeptide, wherein the refolding buffer comprises (i) potassium phosphate or sodium phosphate at a concentration of 25 mM to 150 mM with a pH of 5.8 to 8.0, (ii) guanidine-HCl at a concentration of 0.3 M to 2 M, (iii) L-Arginine at a concentration of 0.25 M to 1 M, (iv) Tween-80 at a concentration of 0.05% to 1%, and (v) oxidized glutathione at a concentration of 1 mM to 4 mM and reduced glutathione at a concentration of 0.05 mM to 0.8 mM, wherein the ratio of oxidized to reduced glutathione is from 5;
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1. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36)
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Specification