Browse > Article

Solid-Phase Refolding Technology in Recombinant Proteins Recovery: Application Examples to Various Biopharmaceutical Proteins  

Kim, Min Young (Bioprocessing Research Laboratory, Department of Chemical Engineering, Hanyang University)
Suh, Chang Woo (Biotechnology Research Center, Daewoong Corp.)
Kim, Chang Sung (Dupont Korea Ltd.)
Jo, Tae Hoon (Department of Chem. Eng., Univ. of Southern California)
Park, Sang Joong (CKD Bio Research Institute, CKD Bio Corp.)
Choi, Won Chan (Quality Control Team, Green Cross Plasma Derivatives, Corp.)
Lee, Eun Kyu (Bioprocessing Research Laboratory, Department of Chemical Engineering, Hanyang University)
Publication Information
Korean Chemical Engineering Research / v.43, no.2, 2005 , pp. 187-201 More about this Journal
Abstract
Bioprocessing technologies utilizing 'biorecognition' between a solid matrix and a protein is being widely experimented as a means to replacing the conventional, solution-based technology. Frequently the matrices are chromatographic resins with specific functional groups exposed outside. Since the reactions of and interactions with the proteins occur as they are attached to the solid matrix, this 'solid-phase' processing has distinct advantages over the solution-phase technology. Solid-phase refolding of inclusion body proteins uses ion exchange resins to adsorb denaturant-dissolved inclusion body. As the denaturant is slowly removed from the micromoiety around the protein, it is refolded into a native, three-dimensional structure. Once the refolding is complete, the folded protein can be eluted by a conventional elution technique such as the salt-gradient. This concept was successfully extended to 'EBA (expanded bed adsorption)-mediated refolding,' in which the denaturant-dissolved inclusion body in whole cell homogenate is adsorbed to a Streamline resin while cell debris and other impurity proteins are removed by the EBA action. The adsorbed protein follows the same refolding steps. This solid-phase refolding process shows the potential to improve the refolding yield, reduce the number of processing steps and the processing volume and time, and thus improve the overall process economics significantly. In this paper, the experimental results of the solid-phase refolding technology applied to several biopharmaceutical proteins of various types are presented.
Keywords
Solid-phase Refolding; Inclusion Body; Biopharmaceutics; Packed Bed Adsorption Chromatography; Expanded Bed Adsorption Chromatography;
Citations & Related Records
Times Cited By KSCI : 1  (Citation Analysis)
연도 인용수 순위
1 Schonor, E. G., Ellis, L. F. and Shonor, B. E., 'Isolation and Purification of Protein Granules from E. coli cells Overproducing Bovine Growth Hormone,' Bio/Technol., 3, 151-154(1985)   DOI
2 Bowden, G. A., Paredes, A. M. and Georgiou, G., 'Structure and Morphology of Protein Inclusion Body in Escherichia coli,' Bio/ Technol., 9, 725-734(1991)   DOI
3 Bam, N. B., Cleland, J. L. and Randolph, T. W., 'Molten Globule Intermediate of Recombinant Human Growth Hormone: Stabilization with Surfactants,' Biotechnol. Prog., 12(6), 801-809(1996)   DOI   ScienceOn
4 Cleland, J. L. and Wang, D. I. C., 'Cosolvent Assisted Protein Refolding,' Bio/Technol., 8, 274-278(1990)
5 Lee, E. K., Cho, T. H. and Suh, C. W., 'In vitro Refolding of Inclusion Body Proteins Directly from E. coli Homogenate in Expanded Bed Adsorption Chromatography,' Korean J. Biotechnol. Bioeng., 16(2), 146-152(2001)   과학기술학회마을
6 Lali, A., Kaul, R., Yu, I. and Mattiason, B., 'Purification of LLactate Dehydrogenase from Crude Homogenate of Porcine Muscle by Expanded Bed Affinity Chromatography (EBAC),' Isolation and Purification, 2, 289-300(1997)
7 Chang, Y. K. and Chase, H. A., 'Expanded Bed Adsorption for the Direct Extraction of Proteins,' Separation for Biotechnology, 3, 106-112(1994)
8 Angles-Cano, E. and Rojas, G., 'Apolipoprotein (a): Structure- Function Relationship at the Lysine-Binding Site and Plasminogen Activator Cleavage Site,' Biol. Chem., 383, 93-99(2002)   DOI   ScienceOn
9 Trieu, V. N. and Uckun, F. M., 'Apolipoprotein (a), a Link Between Atherosclerosis and Tumor Angiogenesis, ' Biochem. Biophy. Res. Comm., 257(3), 714-718(1999)   DOI   ScienceOn
10 Badcoe, I., Smith, C., Wood, S., Halsall, D., Holbrook, J., Lund, P. and Clarke, A., 'Binding of a Chaperonin to the Refolding Intermediates of Lactate Dehydrogenase,' Biochemistry, 30(38), 9195- 9200(1991)   DOI
11 Kiefhaber, T., Rudolph, R., Kohler, H.-H. and Buchner, J., 'Protein Aggregation in vitro and in vivo: A Quantitative Model of the Kinetic Competition Between Folding and Aggregation,' Bio/Technol., 9, 825-829(1991)   DOI
12 Hirata, Y. and Orth, A. N., 'Epidermal Growth Factor (Urogastrone) in Human Fluids: Size Heterogeneity,' J. Clin. Endocrinol. Metab., 48, 673-679(1979)   DOI
13 Smith, M. P., Bulmer, M., Hjorth, R. and Titchener-Hooker, N. J., 'A Comparative Engineering Study of the Use of Expanded Bed and Packed Bed Routes for the Recovery of Labile Proteins from Crude Feedstocks,' Proceedings of 5th World Congress of Chemical Engineering, 2, 565-570(1996)
14 Martin, J., Langer, Y., Boteva, R., Schramal, A., Horwich, A. and Hartl, F., 'Chaperionin-Mediated Protein Folding at the Surface of groEL Through a 'Molten Globule'-Like Intermediate,' Nature, 352, 36-42(1991)   ScienceOn
15 Cho, T. H., Ahn, S. J. and Lee, E. K., 'Refolding of Protein Inclusion Bodies Directly from E. coli Homogenate Using Expanded bed Adsorption Chromatography,' Bioseparation, 10, 189-196(2002)   DOI   ScienceOn
16 Zettlmeissl, G., Rudolph, R. and Jaenicke, R., 'Reconstitution of Lactic Dehydrogenase. Noncovalent Aggregation vs. Reactivation,' Biochemistry, 18(25), 5567-5571(1979)   DOI   ScienceOn
17 Cao, Y., Ji, R. W., Davidson, D., Schaller, J., Marti, D., Sohndel, S., McCance, S. G., O'Reilly, M. S., Llinas, M. and Folkman J., 'Kringle Domanis of Human Angiostatin,' J. Biol. Chem., 271(46), 29461-29467(1996)   DOI   ScienceOn
18 Cleland, J. L., Hedgepeth, C. and Wang, D. I. C., 'Polyethylene Glycol Enhanced Refolding of Bovine Carbonic Anhydrase B,' J. Biol. Chem., 267(19), 13327-13334(1992)
19 Tice, P. A., Mazsaroff, I., Line, N. T. and Reginer, F. E., 'Effect of Large Sample Loads on Column Lifetime in Preparative-Scale Liquid Chromatography,' J. chromatography, 410, 43-51(1987)   DOI   ScienceOn
20 Buchner, J. and Rudolph, R., 'Renaturation, Purification and Characterization of Recombinant Fab-Fragments Produced in Escherichia coli,' Bio/Technology, 9, 157-162(1991)   DOI
21 Kim, Y. H., 'Direct Quantification of $Interferon-{\alpha}$ Inclusion body in Recombinant E. coli. Fermentor Beer Using Reversed-phase HPLC,' M.S. Thesis, Dept. of Chem. Eng., Hanyang Univ., Seoul, Korea(1995)
22 Lee, Y. S., 'Fermentation and Purification Processes for Fusion Protein of Human Epidermal Growth Factor and Angiogenin from Recombinant E. coli,' M.S. Thesis. Hanyang University, Dept. of Chemical Engineering, Korea(1999)
23 Creighton, T. E., 'Process for the Production of a Protein,' US Patent No. 4,977,248(1990)
24 Psarras, K., Ueda, M., Yamamura, T., Ozawa, S., Kitajima, M., Aiso, S., Komatsu, S. and Seno, M., 'Human Pancreatic RNase1- Human Epidermal Growth Factor Fusion; an Entirely Human 'Immunotoxin Analog' with Cytotoxic Properties Against Squamous Cell Carcinomas', Protein Eng., 11(12), 1285-1292(1998)   DOI
25 Park, S. J., Kang, R., Suh, C. W., Chai, Y. G., Kwon, O. B., Park, S. K. and Lee, E. K., 'Solid-Phase Refolding of Poly-Lysine Tagged Fusion Protein of hEGF and Angiogenin,' Biotechnol. Bioprocess Eng., 7(1), 1-5(2002)   DOI   ScienceOn
26 Yoon, J. M., Han, S. H., Kown, O. B., Kim, S. H., Park, M. H. and Kim, B. K., 'Cloning and Cytotoxicity of Fusion Proteins of EGF and Angiogenin,' Life Science, 16, 1435-1445(1999)
27 Shapiro, R., Riordan, J. F. and Vallee, B. L., 'Characteristic Ribonucleolytic Activity of Human Angiogenin,' Biochemistry, 25(12), 3527-3532(1986)   DOI   ScienceOn
28 Thommes, J., Bader, A., Halfar, M., Karau, A. and Kula, M-R., 'Isolation of Monoclonal Antibodies from cell Containing Hybridoma Broth Using a Protein A Coated Adsorbent in Expaneded beds,' J. Chromatography, 752, 111-122(1996)   DOI   ScienceOn
29 Defellippis, M. R., Alter, L. A., Pekar, A. H., Havel, H. A. and Brems D. N., 'Evidence for a Self-Associating Equilibrium Intermediate During Folding of Human Growth Hormone,' Biochemistry, 32(6), 1555-1562(1993)   DOI   ScienceOn
30 Ellman, G. L., 'Tissue Sulfhydryl Groups,' Arch. Biochem. Biophys., 82(1), 70-77(1959)   DOI   ScienceOn
31 Hermann, R., 'Protein Folding, in EPO Applied Technology Series,' 12(7), European Patent Office, Netherlands(1993)
32 Wu, Y. N., Saxena, S. K., Ardelt, W., Gadina, M., Mikulshki, S., Lorenzo, C. D., Alessio, G. D. and Youle, J., 'A Study of the Intracellular Routing of Cytotoxic Ribonucleases,' J. Biol. Chem., 270(29), 17476-17481(1995)   DOI   ScienceOn
33 Soncin, F., Strydom, D. J. and Shapiro, R., 'Interaction of Heparin with Human Angiogenin,' J. Biol. Chem., 272(15), 9818-9824(1997)   DOI   ScienceOn
34 Gekko, K. and Timasheff, S. N., 'Mechanism of Protein Stabilization by Glycerol: Preferential Hydration in Glycerol-Water Mixtures,' Biochemisty, 20(16), 4667-4676(1981)   DOI   ScienceOn
35 MacDonald, N. J., Murad, A. C., Fogler, W. E., Lu, Y. and Sim, K. L., 'The Tumor-Suppressing Activity of Angiostatin Protein Residues Within Kringles 1 to 3,' Biochemical and Biophys. Res. Commu., 264(2), 469-477(1999)   DOI   ScienceOn
36 Cleland, J. L. and Randolph, T. W., 'Mechanism of Polyethylene Glycol Interaction with the Molten Globule Intermediate of Bovine Carbonic Anhydrase B,' J. Biol. Chem., 267(5), 3147- 3153(1992)
37 Buchner, J. and Rudolph, R., 'Renaturation, Purification and Characterization of Recombinant Fab-Fragments Produced in Escherichia coli,' Bio/Technol., 9, 157-162(1991)   DOI
38 Lowe, E. G., Rhind, S. K., Sugrue, R. and Marston, F. A. O., 'Solubilization, Refolding and Purification of Eukaryotic Proteins Expressed in E. coli, Protein Purification: Micro to Macro,' 429-442, Alan R. Liss, Inc.(1987)
39 Fischer, B., Summer, I. and Goodenough, P., 'Isolation, Renaturation, and Formation of Disulfide Bonds of Eukaryotic Proteins Expressed in Escherichia coli as Inclusion Bodies,' Biotech. Bioeng., 41(1), 3-13(1993)
40 Stempfer, G., Neugebauer, B. H. and Rudolph, R., 'Improved Refolding of Immobilized Fusion Protein,' Nat. Biotechnol., 14(3), 329-334(1996)   DOI   ScienceOn
41 Jang, J. H., Kim, J. S., Park, E. J., Yeum, J. S. and Jung, S. I., Korean Patent No. 10-2002-7003512(2002)
42 Tang, W., Sun, Z. Y., Pannell, R., Gurewich, V. and Liu, J. N., 'An Efficient System for Production of Recombinant Urokinasetype Plasminogen Activator,' Protein Exp. Purif., 11(3), 279-283(1997)   DOI   ScienceOn
43 Folkman, J. and Shing, Y., 'Angiogenesis', J. Biol. Chem., 267(16), 10931-10934(1992)
44 Kim, C. S. and Lee, E. K., 'Effect of Operating Parameters On in Vitro Renaturation of a Fusion Protein of Human Growth Hormone and Glutathione S Transferase from Inclusion Body,' Process Biochemistry, 36, 111-117(2000)
45 Carmeliet, P. and Jain, R. K., 'Angiogenesis in Cancer and Other Diseases,' Nature, 407, 249-257(2000)   ScienceOn
46 Feuser, J., Walter, J., Kula, M. R. and Thommes, J., 'Cell/ Adsorbed Interactions in Expanded Bed Adsorption of Proteins,' Bioseparation, 8, 99-109(1999)   DOI   ScienceOn
47 Cleland, J. L. and Wang, D. I. C., 'Refolding and Aggregation of Bovine Anhydrase B: Quasi-Elastic Light Scattering Analysis,' Biochemisty, 29(50), 11072-11078(1990)   DOI   ScienceOn
48 Brems, D. N., 'Solubility of Different Folding Conformers of Bovine Growth Hormone,' Biochemistry, 27(12), 4541-4546(1988)   DOI