Chemistry: natural resins or derivatives; peptides or proteins; – Proteins – i.e. – more than 100 amino acid residues – Hormones – e.g. – prolactin – thymosin – growth factors – etc.
Patent
1994-10-11
1998-09-15
Ulm, John
Chemistry: natural resins or derivatives; peptides or proteins;
Proteins, i.e., more than 100 amino acid residues
Hormones, e.g., prolactin, thymosin, growth factors, etc.
530350, 530303, 530418, 530419, 530420, 530422, 530423, 530424, 435 694, 4351723, 4353201, 4352523, C07K 14475, A61K 3822
Patent
active
058080066
DESCRIPTION:
BRIEF SUMMARY
BACKGROUND OF THE INVENTION
1. Field of the Invention
This invention relates to special buffer solutions and their use for refolding polypeptides.
2. Description of Related Art
For commercial production of many polypeptides and proteins, recombinant DNA techniques have become the method of choice because of the large quantities that can be produced in bacteria and other host cells. Manufacturing recombinant protein involves transfecting or transforming host cells with DNA encoding the desired exogenous protein and growing the cells under conditions favoring expression of the recombinant protein. E. coli and yeast are favored as hosts because they can be made to produce recombinant proteins at high titers.
Numerous U.S. patents on general bacterial expression of recombinant-DNA-encoded proteins exist, including U.S. Pat. No. 4,565,785 on a recombinant DNA molecule comprising a bacterial gene for an extracellular or periplasmic carrier protein and non-bacterial gene; 4,673,641 on coproduction of a foreign polypeptide with an aggregate-forming polypeptide; 4,738,921 on an expression vector with a trp promoter/operator and trp LE fusion with a polypeptide such as insulin-like growth factor (IGF-I); 4,795,706 on expression control sequences to include with a foreign protein; and 4,710,473 on specific circular DNA plasmids such as those encoding IGF-I.
Under some conditions, certain heterologous proteins expressed in large quantities from bacterial hosts are precipitated within the cells in dense aggregates, recognized as bright spots visible within the enclosure of the cells under a phase-contrast microscope. These aggregates of precipitated proteins are referred to as "refractile bodies," and constitute a significant portion of the total cell protein. Brems et al., Biochemistry, 24: 7662 (1985). On the other hand, the aggregates of protein may not be visible under the phase contrast microscope, and the term "inclusion body" is often used to refer to the aggregates of protein whether visible or not under the phase-contrast microscope.
It has been found that the soluble proportion of high-level expressed protein in E. coli has been dramatically increased by lowering the temperature of fermentation to below 30.degree. C. A considerable fraction of various foreign proteins, i.e., human interferon-alpha (IFN-.alpha.2), Noteborn, Bio/Technology, 6: 291-294 (1988)! and human IFN-.beta. solution. This procedure represents an alternative to renaturation of proteins recovered from refractile bodies, but requires an expression system that is efficiently induced at temperatures below 30.degree. C. The procedure is therefore not effective for all proteins.
For general review articles on refractile bodies, see Marston, supra; Mitraki and King, Bio/Technology, 7: 690 (1989); Marston and Hartley, Methods in Enzymol., 182: 264-276 (1990); Wetzel, "Protein Aggregation In Vivo: Bacterial Inclusion Bodies and Mammalian Amyloid, " in Stability of Protein Pharmaceuticals: In Vivo Pathways of Degradation and Strategies for Protein Stabilization, Ahern and Manning (eds.) (Plenum Press, 1991); and Wetzel, "Enhanced Folding and Stabilization of Proteins by Suppression of Aggregation In Vitro and In Vivi, " in Protein Engineering--A Practical Approach, Rees, A. R. et. al. (eds.) (IRL Press at Oxford University Press, Oxford, 1991).
Recovery of the protein from these bodies has presented numerous problems, such as how to separate the protein encased within the cell from the cellular material and proteins harboring it, and how to recover the inclusion body protein in biologically active form. The recovered proteins are often predominantly biologically inactive because they are folded into a three-dimensional conformation different from that of active protein. For example, misfolded IGF-I with different disulfide bond pairs than found in native IGF-I has significantly reduced biological activity. Raschdorf et al., Biomedical and Environmental Mass Spectroscopy, 16: 3-8 (1988). Misfolding occurs either in the cell during fermentation or du
REFERENCES:
patent: 4511502 (1985-04-01), Builder et al.
patent: 4511503 (1985-04-01), Olson et al.
patent: 4512922 (1985-04-01), Jones et al.
patent: 4518256 (1985-05-01), Schwartz
patent: 4518526 (1985-05-01), Olson
patent: 4565785 (1986-01-01), Gilbert
patent: 4572798 (1986-02-01), Koths et al.
patent: 4620948 (1986-11-01), Builder et al.
patent: 4652630 (1987-03-01), Bentle et al.
patent: 4673641 (1987-06-01), George et al.
patent: 4705848 (1987-11-01), Yang et al.
patent: 4710473 (1987-12-01), Morris
patent: 4738921 (1988-04-01), Bellagaje et al.
patent: 4795706 (1989-01-01), Hsiung et al.
patent: 4923967 (1990-05-01), Bobbitt et al.
patent: 4985544 (1991-01-01), Yokoo et al.
patent: 5019500 (1991-05-01), Ueda et al.
patent: 5028531 (1991-07-01), Ueda et al.
patent: 5158875 (1992-10-01), Miller et al.
patent: 5191063 (1993-03-01), Inouye et al.
patent: 5288931 (1994-02-01), Chang et al.
Blundell et al., "Tertiary structures, receptor binding, and antigenicity of insulinlike growth factors" Federation Proc. 42:2592-2597 (1983).
Boss et al., "Assembly of fucntional antibodies from immunoglobulin heavy and light chains synthesized in E. coli" Nucleic Acids Research 12(9):3791-3806 (1984).
Bowden et al., "Structure and Morphology of Protein Inclusion Bodies in Escherichia coli" Bio/Technology 9:725-730 (Aug. 1991).
Brems D. et al., "Equilibrium denaturation of pituitary- and recombinant-derived bovine growth hormone" Biochemistry 24(26):7662-7668 (1985).
Brems et al., "Altering the association properties of insulin by amino acid replacement" Protein Engineering 5(6):527-533 (1992).
Brems et al., "Equilibrium Denaturation of Insulin and Proinsulin" Biochemistry 29:9289-9293 (1990).
Brems et al., "Improved insulin stability through amino acid substitution" Protein Engineering 5(6):519-525 (1992).
Bryant et al., "Detection of an Equilibrium Intermediate in the Folding of a Monomeric Insulin Analog" Biochemistry 31(25):5692-5698 (1992).
Cabilly et al., "Generation of antibody activity from immunoglobulin polypeptide chains produced in Escherichia coli" Proc. Natl. Acad. Sci. USA 81:3273-3277 (1984).
Cleland, "Impact of protein folding on biotechnology" Protein Folding: In Vivo and In Vitro (ACS Symposium Series 526), Cleland (ed.), Washington, D.C.:American Chemical Society p. 6 (1993).
Cleland et al., "Refolding and Aggregation of Bovine Carbonic Anhydrase B: Quasi-Elastic Light Scattering Analysis" Biochemistry 29:11072-11078 (1990).
Cooke et al., "Solution Structure of Human Insulin-Like Growth Factor 1: A Nuclear Magnetic Resonance and Restrained Molecular Dynamics Study" Biochemistry 30:5484-5491 (1991).
Creighton, "Disulfide Bond Formation in Proteins" Methods in Enzymology 107:305-329 (1984).
Creighton, "Role of the Environment in the Refolding of Reduced Pancreatic Trypsin Inhibitor" J. Mol. Biol. 144:521-550 (1980).
Frank et al., "The Production of Human Proinsulin and its Transformation to Human Insulin and C-Peptide" Peptides: Synthesis, Structure, Function, Rich and Gross pp. 729-738 (1981).
Fu and Freire, "On the origin of the enthalpy and entropy convergence temperatures in protein folding" Proc. Natl. Acad. Sci. USA 89:9335-9338 (1992).
Funakoshi et al., "Isolation of a Tumor-Derived 186-Residue Peptide Amide Related to Human Chromogranin A and its in vitro Conversion to Human Pancreastatin-48" Recent Bioactive Peptides and Biology pp. 512-514 (1989).
Furman et al., "Recombinant Human Insulin-Like Growth Factor II Expressed in Escherichia coli" Bio/Technology 5:1047-1051 (1987).
George et al., "High-Level Expression in Escherichia coli of Biology Active Bovine Growth Hormone" DNA 4:273-281 (1985).
Gill et al., "Recombinant Chicken and Bovine Growth Hormones Accelarate Growth in Aquacultured Juvenile Pacific Salmon Oncorhynchus Kisutch" Bio/Technology 3:643-646 (1985).
Gold, "Purification of Biosynthetic Human Relaxin A-Chain and B-Chain and Scaleup of the Chain Combination Reaction" Protein Folding and Recovering Symposium (1992).
Green et al.,
Builder Stuart
Hart Roger
Lester Philip
Reifsnyder David
Genentech Inc.
Hasak Janet E.
Mertz Prema
Ulm John
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