Method for refolding enzymes

Details Method for refolding enzymes Method for refolding enzymes

2007-03-06

2007-03-06

Nashed, Nashaat T. (Department: 1656)

Chemistry: molecular biology and microbiology

Enzyme , proenzyme; compositions thereof; process for...

Hydrolase

C435S226000

Reexamination Certificate

active

10230677

ABSTRACT:
The invention provides methods for efficient recombinant expression, refolding, and purification of Beta-site APP cleaving enzyme (BACE) polypeptides. In various aspects, the method includes the steps of expressing a recombinant construct in bacteria, dissolving inclusion bodies with a denaturant at high pH in the presence of a reducing agent, diluting the solubilized BACE polypeptide in an aqueous solution at a temperature of about 1° C. to 15° C., and incubating the diluted sample at a temperature of about 4° C. to 15° C. until the recombinant BACE polypeptide folds into an active enzyme.

REFERENCES:
patent: 5744346 (1998-04-01), Chrysler et al.
patent: 6319689 (2001-11-01), Powell et al.
patent: 6323326 (2001-11-01), Dorin et al.
patent: 6545127 (2003-04-01), Tang et al.
patent: 6583268 (2003-06-01), Lin
patent: 9822597 (1998-05-01), None
patent: 0017369 (2000-03-01), None
patent: 0047618 (2000-08-01), None
patent: 0100663 (2001-01-01), None
patent: 0123533 (2001-04-01), None
Yan, Riquang, et al;Membrane-anchored aspartyl protease with Alzheimer's diseaseβsecretase activity; Nature, vol. 402, Dec. 2, 1999.
Vassar, Robert, et al; β-Secretase Cleavage of Alzheimer's Amyloid Precursor Protein by the Transmembrane Aspartic Protease BACE; Science, vol. 286, Oct. 22, 1999, pp. 735-741.
Sinha, Sukanto, et al;Purification and cloning of amyloid precursor prtein β -secretase from human brain; Nature, vol. 402, (6761):537-540.
Mallender, William;Characterization of Recombinant, Soluble β-Secretase from an Insect Cell Expression System; Molecular Pharmocology; 59:619-626, 2001.
Benjannet, Suzanne, et al;Post-translation Processingof β-Secretase (β-Amyloid-converting Enzyme) and its Ectodomain Shedding; The Journal of Biological Chemistry, vol. 276, No. 14, Apr. 6, 2001; 10879-10887.
Capell, Anja et al.Maturation of Pro-peptide Cleavage of β-Secretase; The Journal of Biological Chemistry, vol. 275, No. 40, Oct. 6, 2000; pp. 30849-30854.
Charlwood, Joanne, et al; Characterization of the Blycosylation profiles of Alzheimer's β-Secretase Protein Asp-2 Expressed ina variety of Cells Lines; The Journal of Biological Chemistry, vol. 276, No. 20, May 18, 2001; p. 16739-16748.
Ermolieff Jacques, et al; Proteolytic Activation of Recombinant Pro-memapsin 2 (Pro-β-secetase) Studies with New Fluorogenic Substance; Biochemistry 2000, 39, 12450-12456.
Haniu, Mitsuru et al;Characterization of Alzheimer's β-Secretase Protein BACE; The Journal of Biological Chemistry; vol. 275, No. 28, Jul. 14, 2000: pp. 21099-21106.
Hussain, Ishrut, et al;Identification of a Novel Aspartic Protease(Asp 2)as β-Scretase; Molecular and Cellular Neuroscience; 14, 419-427 (1999).
Khan, Amir, et al;Molecular mechanisms for the convesion of zymogens to active proteolytic enzymes; Protein Science (1998) 7:815-836.
Lin, Xinli, et al;Rearranging the domains of pepsinogen; (1995), 4: 159-166.
Lin, Xin-Li, et al;Synthesis, Purification, and Active Site Mutagenesis of Recombinant Procine Pepsinogen; The Journal of Biological Chemistry; vol. 264, No. 8, Mar. 15, 1989; pp. 4482-4489.
Lin, Xinli, et al;Human aspartic proteas emeapsin 2 cleaves theβ-amyloid precursor protein; PNAS, Feb. 15, 2000; vol. 97, pp. 1456-1460.
Mildner, Ana, et al;Production of Chemokines CTAPIII and NAP/2 by Digestion of Recombinant Ubiquitin-CTAPIII with Yeast Ubiquitin C-Terminal Hydrolase and Human Immunodeficiency Virus Protease; Protein Expression and Purification vol. 16, 347-354 (1999).
Selkoe, D.J.,Cell Biology of theβ-Amyloid Precursor Protein and the Genetics of alzheimer's Disease; Cold Spring Harbor Symposia on Quantative Biology, vol. LXI, 1996, pp. 587-596.
Selkoe, Dennis J.,;translating cell biology into therapeutic advances in Alzheimer's disease; Nature, vol. 399, Jun. 24, 199 pp. A23-A31.
Thinakaran, Gopal, et al;Metabolis of the “Swedish” Amyloid Precursor Protein Variant in Neur2a(N2a)Cells; the Journal of Biological Chemistry; vol. 271, No. 16, Apr. 19, 1996; pp. 9390-3997.
Hong, L. et al; Structure of the protease domain of memapsin 2 (beta-secretase) complexed with inhibitor; Science 2000; 290(5489), 150-3.
Heinrikson, R.L., et al; The Biochemistry and molecular biology of recombinant rennin and proorenin; Hypertension: Pathophysiology, Diagnosis and Management 1990; chapter 74:1179-1196.
Inagami, T., et al: Biomed Res. 1980; 1:456-475.
Schechter, Israel, et al;On the Size of the Active site in Proteases, I Papain, Biochemical and Biophysical Research Communications, vol. 27, No. 2, 1967; pp. 157-162.
Selkoe, Dennis, J.;Alzheimer's Disease: Genes, Proteins and Therapy, Physiological Review, vol. 87, No. 2, Apr. 2001, pp. 741-766.

Affiliated with

Also associated with

Rating

Method for refolding enzymes does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Method for refolding enzymes, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Method for refolding enzymes will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-3781322