Refolding of recombinant proteins

Current Opinion in Biotechnology

Volumn 9, Issue 2, 1998, Pages 157-163

  De Bernardez Clark, Eliana ^a  

^a Tufts University   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

CELL PROTEIN; CHAPERONE; FOLDASE; RECOMBINANT PROTEIN; UNCLASSIFIED DRUG;

BACTERIUM; CELL INCLUSION; NONHUMAN; PRIORITY JOURNAL; PROTEIN AGGREGATION; PROTEIN EXPRESSION; PROTEIN FOLDING; PROTEIN SYNTHESIS; RENATURATION; REVIEW;

EID: 0031950560     PISSN: 09581669     EISSN: None     Source Type: Journal    
DOI: 10.1016/S0958-1669(98)80109-2     Document Type: Article

References (44)

1. Clarke AR, Waltho JP. Protein folding pathways and intermediates. Curr Opin Biotechnol. 8:1997;400-405.
2. Herman R. Protein Folding. 1993;European Patent Office, Munich. [EPO Applied Technology Series vol 12.].
3. Rudolph R, Böhm G, Lilie H, Jaenicke R. Folding proteins. of special interest Creighton TE. edn 2 Protein Function. A Practical Approach. 1997;57-99 IRL Press, New York, This chapter covers relevant topics and techniques in protein unfolding/folding. It contains protocols for cell lysis, isolation of inclusion bodies, and renaturation of solubilized proteins with special emphasis on the renaturation of disulphide bonded proteins. It also covers monitoring of protein folding, kinetics and equilibrium considerations, folding and association of oligomeric proteins, the effects of ligands and the effect of auxiliary proteins assisting protein folding.
4. Cardamone M, Puri NK, Brandon MR. Comparing the refolding and reoxidation of recombinant porcine growth hormone from a urea denatured state and from Escherichia coli inclusion bodies. Biochemistry. 34:1995;5773-5794.
5. Cowley DJ, Mackin RB. Expression, purification and characterization of recombinant human proinsulin. of special interest FEBS Lett. 402:1997;124-130 E. coli derived recombinant human proinsulin produced in inclusion bodies was solubilized with 70% formic acid and cleaved with cyanogen bromide. After solvent evaporation and freeze-drying, the lyophilized protein was dissolved in 7 M urea and subjected to oxidative sulphitolysis. The sulphonated material was purified by anion exchange, and refolded at pH 10.5 in the presence of air and β-mercaptoethanol. The refolded proinsulin contained the correct disulphide bond pattern.
6. Burgess RR. Purification of overproduced Escherichia coli RNA polymerase σ factors by solubilizing inclusion bodies and refolding from sarkosyl. Methods Enzymol. 273:1996;145-149.
7. Kurucz I, Titus JA, Jost CA, Segal DM. Correct disulphide pairing and efficient refolding of detergent-solubilized single-chain Fv proteins from bacterial inclusion bodies. Mol Immunol. 12:1995;1443-1452.
8. Stöckel J, Döring K, Malotka J, Jähnig F, Dornmair K. Pathway of detergent-mediated and peptide ligand-mediated refolding of heterodimer class II major histocompatibility complex (MHC) molecules. of special interest Eur J Biochem. 248:1997;684-691 An in depth discussion of the mechanism of detergent-mediated protein folding of a heterodimeric disulphide bonded protein with four domains. The contributions of detergent headgroup and aliphatic tail to the stabilization of folding intermediates are dissected. Optimal detergent concentration decreases with increasing critical micelle concentration. Formation of secondary structure occurs early in the folding pathway when the denaturing detergent SDS is replaced by a mild detergent. Tertiary structure formation and heterodimer association occurs later in the folding pathway concomitantly with disulphide bond formation.
9. Maachupalli-Reddy J, Kelley BD, De Bernardez Clark E. Effect of inclusion body contaminants on the oxidative renaturation of hen egg white lysozyme. of special interest Biotechnol Prog. 13:1997;144-150 This paper shows that non-proteinaceous contaminants, such as plasmid DNA, ribosomal RNA, and lipopolysaccharides, have little effect on protein renaturation. Phospholipids improve folding yields by about 15%. Proteinaceous contaminants, on the other hand, can have a significant detrimental effect on folding yields by causing co-aggregation of the protein of interest. The paper also shows that contaminants affect the overall rate of the aggregation reaction without affecting the folding rate.
10. Ahn JH, Lee YP, Rhee JS. Investigation of refolding condition for Pseudomonas fluorescens lipase by response surface methodology. J Biotechnol. 54:1997;151-160.
11. Simmons T, Newhouse YM, Arnold KS, Innerarity TL, Weisgraber KH. Human low density lipoprotein receptor fragment. Successful refolding of a functionally active ligand-binding domain produced in Escherichia coli. J Biol Chem. 272:1997;25531-25536.
12. Negro A, Onisto M, Grassato L, Caenazzo C, Garbisa S. Recombinant human TIMP-3 from Escherichia coli: synthesis, refolding, physico-chemical and functional insights. Protein Eng. 10:1997;593-599.
13. 15N] enriched guinea pig [Val90]-α-lactalbumin. Protein Eng. 10:1997;455-462.
14. Buchner J, Rudolph R. Renaturation, purification and characterization of recombinant Fab fragments produced in Escherichia coli. Biotechnol. 9:1991;157-162.
15. Hamaker KH, Liu J, Seely RJ, Ladisch CM, Ladisch MR. Chromatography for rapid buffer exchange and refolding of secretory leukocyte protease inhibitor. Biotechnol Prog. 12:1996;184-189.
16. Batas B, Jones HR, Chaudhuri JB. Studies of the hydrodynamic volume changes that occur during refolding of lysozyme using size-exclusion chromatography. J Chromatog A. 766:1997;109-119.
17. Batas B, Chaudhuri JB. Protein refolding at high concentration using size-exclusion chromatography. Biotechnol Bioeng. 50:1996;16-23.
18. Qiu H, Wen D, Belanger A, Bunn HF. Over-expression and refolding of biologically active human erythropoietin from E. coli. Protein Eng. 10:1997;33. (suppl).
19. Stempfer G, Rudolph R. Improved refolding of a matrix-bound fusion protein. Ann N Y Acad Sci. 782:1996;506-512.
20. Stempfer G, Höll-Neugebauer B, Rudolph R. Improved refolding of an immobilized fusion protein. of special interest Nat Biotechnol. 14:1996;329-334 The authors demonstrate a strategy to prevent aggregation upon folding by physically isolating protein molecules from each other through immobilizing onto a solid support. In this technique, the protein of interest is produced as a fusion protein with either an amino- or a carboxy-terminal hexa-arginine peptide. The denatured protein is attached to a solid support containing polyanionic groups and renaturation on the immobilized protein is performed by removal of the denaturant. The effect of ionic strength, addition of ethylene glycol and support material are thoroughly investigated.
21. Rudolph R, Lilie H. In vitro folding of inclusion body proteins. of special interest FASEB J. 10:1996;49-56 An excellent review with 80 references detailing key aspects of expression and refolding of inclusion body proteins.
22. Hevehan D, De Bernardez Clark E. Oxidative renaturation of lysozyme at high concentrations. of special interest Biotechnol Bioeng. 54:1997;221-230 This paper shows that it is possible to refold a disulphide-bonded protein with very high yields at high protein concentrations (up to 5 mg/ml) by addition of non-denaturing concentrations of the chaotropic agent guanidinium chloride (GdmCl). Increasing GdmCl concentrations decreased both the rate of the folding and aggregation reactions, with a stronger effect on aggregation. The paper also shows that GdmCl is as efficient as L-arginine in decreasing aggregation while resulting in faster folding rates.
23. Xie Y, Wetlaufer DB. Control of aggregation in protein folding: the temperature-leap tactic. Protein Sci. 5:1996;517-523.
24. Maeda Y, Ueda T, Imoto T. Effective renaturation of denatured and reduced immunoglobulin G in vitro without assistance of chaperone. Protein Eng. 9:1996;95-100.
25. Ptitsyn OB, Bychkova VE, Uversky VN. Kinetic and equilibrium folding intermediates. Philos Trans R Soc Lond B Biol Sci. 348:1995;35-41.
26. Creighton TE. How important is the molten globule for correct protein folding? of special interest Trends Biochem Sci. 22:1997;6-10 In this paper, TE Creighton analyzes experimental observations of the oxidative refolding of α-lactalbumin obtained in his laboratory and by other investigators. He shows that, contrary to what is widely believed, the molten globule has little native-like topology and it could be an off-pathway, non productive species, rather than the key to rapid folding.
27. Yon JM. The specificity of protein aggregation. Nat Biotechnol. 14:1996;1231.
28. Goldberg ME, Rudolph R, Jaenicke R. A kinetic study of the competition between renaturation and aggregation during the refolding of denatured-reduced egg white lysozyme. Biochemistry. 30:1991;2790-2797.
29. Speed MA, Wang DIC, King J. Specific aggregation of partially folded polypeptide chains: the molecular basis of inclusion body composition. of special interest Nat Biotechnol. 14:1996;1283-1287 The authors refolded a mixture of P22 tailspike and coat proteins under conditions which promote aggregation of both proteins. Co-aggregation of the proteins was not observed indicating that aggregation is due to specific interactions. As only soluble multimeric species were analyzed, the possibility of formation of heterogeneous insoluble aggregates cannot be excluded.
30. De Bernardez Clark E, Hevehan D, Szela S, Maachupalli-Reddy J. Oxidative renaturation of hen egg-white lysozyme. Folding vs. aggregation. Biotechnol Prog. 14:1998;47-54.
31. Speed MA, King J, Wang DIC. Polymerization mechanism of polypeptide chain aggregation. Biotechnol Bioeng. 54:1997;333-343.
32. Knappik A, Plückthun A. Engineered turns of a recombinant antibody improve its in vivo folding. Protein Eng. 8:1995;81-89.
33. Nieba L, Honegger A, Krebber C, Plückthun A. Disrupting the hydrophobic patches at the antibody variable/constant domain interface: improved in vivo folding and physical characterization of an engineered scFv fragment. of special interest Protein Eng. 10:1997;435-444 Amino acid mutations away from the antibody fragment binding site resulted in decreased aggregation rates in vitro. Mutations were performed on exposed hydrophobic residues which are adjacent to another exposed hydrophobic residue in order to disrupt hydrophobic patches. The paper also explores the role of these mutations on in vivo folding and aggregation of the resulting scFv fragments.
34. Katzav-Gozansky T, Hanan E, Solomon B. Effect of monoclonal antibodies in preventing carboxypeptidase A aggregation. of special interest Biotechnol Appl Biochem. 23:1996;227-230 This paper shows that certain monoclonal antibodies that bind to the antigen at locations removed from the active site, are capable of promoting proper folding by decreasing aggregation. Thus, monoclonal antibodies can be used as folding additives to prevent aggregation, and as tools to identify sites where protein aggregation may be initiated.
35. Rozema D, Gellman SH. Artificial chaperone-assisted refolding of denatured-reduced lysozyme: modulation of the competition between renaturation and aggregation. of special interest Biochemistry. 35:1996;15760-15771 The artificial chaperone strategy consists of refolding by dilution in the presence of a detergent followed by stripping of the detergent using methyl-β-cyclodextrin. Only ionic detergents are effective in the role of artificial chaperones. Lysozyme is not active in the presence of detergent, and activity is recovered only after the stripping step. Thiol/disulphide reagents can be added either in the dilution step (in the presence of detergent) or in the stripping step. Higher yields are obtained when thiol/disulphide exchange occurs in the presence of detergents.
36. Thomas JG, Ayling A, Baneyx F. Molecular chaperones, folding catalysts, and the recovery of active recombinant proteins from E. coli. To fold or to refold. of special interest Appl Biochem Biotechnol. 66:1997;197-238 A detailed review of the role of chaperones and foldases on in vivo and in vitro protein folding. Includes an extensive list of relevant references.
37. King J. Refolding with a piece of the ring. Nat Biotechnol. 15:1997;514-515.
38. Altamirano M, Golbik R, Zahn R, Bucle AM, Fersht AR. Refolding chromatography with immobilized mini-chaperones. of special interest Proc Natl Acad Sci USA. 94:1997;3576-3578 The authors demonstrate the practical use of the GroEL apical domain (residues 191-376) and the core of the apical domain (residues 191-345) as folding enhancers in the renaturation of several difficult to refold proteins. The mini-chaperones are immobilized on chromatographic resins and chromatography can be conducted in both elution and batch modes. The technique only works with proteins that are known to be GroEL substrates. Its applicability in the oxidative renaturation of proteins is not demonstrated.
39. Rozema D, Gellman SH. Artificial chaperone-assisted refolding of carbonic anhydrase B. J Biol Chem. 271:1996;3478-3487.
40. Couthon F, Clottes E, Vial C. Refolding of SDS- and thermally denatured MM-creatine kinase using cyclodextrins. Biochem Biophys Res Commun. 227:1996;854-860.
41. Wetlaufer DB, Xie Y. Control of aggregation in protein refolding: a variety of surfactants promote renaturation of carbonic anhydrase II. Protein Sci. 4:1995;1535-1543.
42. Maeda Y, Yamada H, Ueda T, Imoto T. Effect of additives on the renaturation of reduced lysozyme in the presence of 4 M urea. of special interest Protein Eng. 9:1996;461-465 Increasing concentrations of additives such as sarcosine, glycerol, ammonium sulphate, glucose and N-acetyl glucosamine resulted in improved refolding rates (except for glycerol) and yields in the oxidative renaturation of lysozyme. Sarcosine was the most effective folding enhancer.
43. Goldberg ME, Expert-Bezanlon N, Vuillard L, Rabilloud T. Non-detergent sulphobetaines: a new class of molecules that facilitate in vitro protein renaturation. Fold Design. 1:1996;21-27.
44. Bam NB, Cleland JL, Randolph TW. Molten globule intermediate of recombinant human growth hormone: stabilization with surfactants. Biotechnol Prog. 12:1996;801-809.