Clinical manufacturing of recombinant human interleukin 15. I. Production cell line development and protein expression in E. coli with stop codon optimization

Biotechnology Progress

Volumn 28, Issue 2, 2012, Pages 497-507

  Vyas, Vinay V ^a   Esposito, Dominic ^a   Sumpter, Terry L ^a   Broadt, Trevor L ^a   Hartley, James ^a   Knapp Iv, George C ^a   Cheng, Wei ^a   Jiang, Man Shiow ^a   Roach, John M ^a   Yang, Xiaoyi ^a   Giardina, Steven L ^a   Mitra, George ^a   Yovandich, Jason L ^a   Creekmore, Stephen P ^a   Waldmann, Thomas A ^b   Zhu, Jianwei ^a  

^a SAIC FREDERICK INC   (United States)

^b NATIONAL CANCER INSTITUTE   (United States)

Author keywords

Clinical manufacturing; E. coli; Interleukin 15; Protein expression; Stop codon

Indexed keywords

ANTITUMOR IMMUNITY; BIOLOGICAL PROPERTIES; CDNA SEQUENCE; CELL LINES; CODING SEQUENCES; CODON OPTIMIZATION; E. COLI; EXPRESSION CONSTRUCTS; HOST CELLS; HUMAN INTERLEUKIN; INCLUSION BODIES; INTERLEUKIN 15; MASS SPECTROMETRY ANALYSIS; NOVEL STRATEGIES; PEPTIDE MAPPING; PRODUCT QUALITY; PRODUCTION CELLS; PROTEIN EXPRESSIONS; SHAKE FLASKS; SIMPLE METHOD; STOP CODON; T CELL ACTIVATION; T CELLS; TRYPTOPHAN RESIDUES;

AMINO ACIDS; CELL CULTURE; CELLS; MANUFACTURE; MASS SPECTROMETRY; OPTIMIZATION; PURIFICATION;

ESCHERICHIA COLI;

INTERLEUKIN 15; RECOMBINANT PROTEIN;

ARTICLE; CELL PROLIFERATION; DRUG EFFECT; ESCHERICHIA COLI; GENE EXPRESSION; GENETICS; HUMAN; METABOLISM; PROTEIN ENGINEERING; STOP CODON;

CELL PROLIFERATION; CODON, TERMINATOR; ESCHERICHIA COLI; GENE EXPRESSION; HUMANS; INTERLEUKIN-15; PROTEIN ENGINEERING; RECOMBINANT PROTEINS;

ESCHERICHIA COLI;

EID: 84859647334     PISSN: 87567938     EISSN: 15206033     Source Type: Journal    
DOI: 10.1002/btpr.746     Document Type: Article

References (32)

1. Grabstein KH, Eisenman J, Shanebeck K, Rauch C, Srinivasan S, Fung V, Beers C, Richardson J, Schoenborn MA, Ahdieh M, Johnson L, Alderson MR, Watson JD, Anderson DM, Giri JG. Cloning of a T-cell growth factor that interacts with the b chain of the interleukin-2 receptor. Science. 1994; 264: 965-968.
2. Burton JD, Bamford RN, Peters C, Grant AJ, Kurys G, Goldman CK, Roessler E, Waldmann TA. A lymphokine, provisionally designated interleukin T and produced by a human adult T-cell leukemia line, stimulates T-cell proliferation and the induction of lymphokine-activated killer cells. Proc Natl Acad Sci USA. 1994; 91: 4935-4939.
3. Waldmann TA. The biology of interleukin-2 and interleukin-15: implications for cancer therapy and vaccine design. Nat Rev Immunol. 2006; 6: 595-601.
4. Klebanoff CA, Finkelstein SE, Surman DR, Lichtman MK, Gattinoni L, Theoret MR, Grewal N, Spiess PJ, Antony PA, Palmer DC, Tagaya Y, Rosenberg SA, Waldmann TA, Restifo NP. IL-15 enhances the in vivo antitumor activity of tumor-reactive CD8+ T cells. Proc Natl Acad Sci USA. 2004; 101: 1969-1974.
5. Teague RM, Sather BD, Sacks JA, Huang MZ, Dossett ML, Morimoto J, Tan X, Sutton SE, Cooke MP, Ohlén C, Greenberg PD. Interleukin-15 rescues tolerant CD8+ T cells for use in adoptive immunotherapy of established tumors. Nat Med. 2006; 12: 335-341.
6. Zhang M, Yao Z, Dubois S, Ju W, Müller JR, Waldmann TA. Interleukin-15 combined with an anti-CD40 antibody provides enhanced therapeutic efficacy for murine models of colon cancer. Proc Natl Acad Sci USA. 2009; 5: 106: 7513-7518.
7. Lugli F, Goldman CK, Perera LP, Smedley J, Pung R, Yovandich JL, Creekmore SP, Waldmann TA, Roederer M. Transient and persistent effects of IL15 on lymphocyte homeostasis in nonhuman primates. Blood. 2010; 116: 3238-3248.
8. Saikh KU, Kissner TL, Nystrom S, Ruthel G, Ulrich RG. Interleukin-15 increases vaccine efficacy through a mechanism linked to dendritic cell maturation and enhanced antibody titers. Clin Vaccine Immunol. 2008; 15: 131-137.
9. d'Ettorre G, Forcina G, Lichtner M, Mengoni F, D'Agostino C, Massetti AP, Mastroianni CM, Vullo V. Interleukin-15 in HIV infection: immunological and virological interactions in antiretroviral-naive and -treated patients. AIDS. 2002; 16: 181-188.
10. Kutzler MA, Robinson TM, Chattergoon MA, Choo DK, Choo AY, Choe PY, Ramanathan MP, Parkinson R, Kudchodkar S, Tamura Y, Sidhu M, Roopchand V, Kim JJ|, Pavlakis PN, Felber BK, Waldmann TA, Boyer JD, Weiner DB. Co-immunization with an optimized IL-15 plasmid results in enhanced function and longevity of CD8 T cells that are partially independent of CD4 T-cell help. J Immunol. 2005; 175: 112-123.
11. Bergamaschi C, Jalah R, Kulkarni V, Rosati M, Zhang GM, Alicea C, Zolotukhin AS, Felber BK, Pavlakis GN. Secretion and biological activity of short signal peptide IL-15 is chaperoned by IL-15 receptor alpha in vivo. J Immunol. 2009; 183: 3064-3072.
12. Cheever MA. Twelve immunotherapy drugs that could cure cancers. Immunol Rev. 2008; 222: 357-368.
13. Jalah R, Rosati M, Kulkarni V, Patel V, Bergamaschi C, Valentin A, Zhang GM, Sidhu MK, Eldridge JH, Weiner DB, Pavlakis GN, Felber BK. Efficient systemic expression of bioactive IL-15 in mice upon delivery of optimized DNA expression plasmids. DNA Cell Biol. 2007; 26: 827-840.
14. Pettit DK, Bonnert TP, Eisenman J, Srinivasan S, Paxton R, Beers C, Lynchi D, Milleri B, Yost J, Grabstein KH, Gombotz WR. Structure-function studies of interleukin 15 using site-specific mutagenesis polyethylene glycol conjugation, and homology modeling. J Biol Chem. 1997; 272: 2312-2318.
15. WHO. WHO reference reagent Interleukin-15 (Human rDNA-derived) NIBSC code:95/554 instruction for use (Version 4.0, Dated 30/01/2008). Available at:
16. Hanick NA, Rickert M, Varani L, Bankovich AJ, Cochran JR, Kim DM, Surh CD, Garcia KC. Elucidation of the interleukin-15 binding site on its alpha receptor by NMR. Biochemistry. 2007; 46: 9453-9461.
17. Ward A, Anderson M, Craggs RI, Maltby J, Grahames C, Davies RA, Finch D, Pattison D, Oakes H, Mallinder PR. E. coli expression and purification of human and cynomolgus IL-15. Protein Expr Purif. 2009; 68: 42-48.
18. Mortier E, Quéméner A, Vusio P, Lorenzen I, Boublik Y, Grötzinger J, Plet A, Jacques Y. Soluble interleukin-15 receptorα (IL-15Rα)-sushi as a selective and potent agonist of IL-15 action through IL-15Rβ/γ. J Biol Chem. 2006; 281: 1612-1619.
19. Xia ZJ, Kong XL, Zhang P. In vivo effect of recombined IL-15/Fc fusion protein on EAU. Sichuan Da Xue Xue Bao Yi Xue Ban. 2008; 39: 944-949.
20. Vyas VV. Clinical manufacturing of recombinant human Interleukin 15. Biotech Progress. To be submitted.
21. Nellis D. Characterization of deamidation of recombinant IL-15. Pharm Res. In press. DOI: 10.1007/s11095-011-0597-0.
22. NCI. A phase I study of intravenous recombinant human IL-15 in adults with refractory metastatic malignant melanoma and metastatic renal cell cancer. 2009. Available at:. Last retrieve date: Nov 29, 2011.
23. Esposito D, Garvey LA, Chakiath CS. Gateway cloning for protein expression. In: Sharon AD, editor. Method in Molecular Biology: High-throughput protein expression and purification. Totowa, NJ: Humana Press; 2009: 31-54.
24. Sambrook J, Russell D. Molecular cloning: A laboratory manual, 3rd ed. Cold Spring Harbor, New York: Cold Spring Harbor Laborator Press; 2001
25. Soman G, Yang X, Jiang H, Giardina S, Vyas V, Mitra G, Yovandich J, Creekmore SP, Waldmann TA, Quinones O, Alvord WG. MTS dye-based colorimetric CTLL-2 cell proliferation assay for product release and stability monitoring of Interleukin-15: Assay qualification, standardization, and statistical analysis. J Immonol Methods. 2009; 348: 83-94.
26. MacBeath G, Kast P. UGA read-through artifacts-when popular gene expression systems need a pATCH. BioTechniques. 1998; 24: 789-794.
27. Geller, AI, Rich, AP. A UGA termination suppression tRNATrp active in rabbit reticulocytes. Nature. 1980; 283: 41-46.
28. Li G, Rice C. The signal for translational read-through of a UGA codon in sindbis virus RNA involves a single cytidine residue immediately downstream of the termination codon. J Virol. 1993; 67: 5062-5067.
29. Hatfield, DL, Smith DWE, Lee BJ, Worland PJ, Oroszlan S. Structure and function of suppressor tRNAs in higher eukaryotes. Crit Rev Biochem Mol Biol. 1990; 25: 71-96.
30. Brown CM, Stockwell PA, Trotman CAN, Tate WP. Sequence analysis suggests that tetra-nucleotides signal the termination of protein synthesis in eukaryotes. Nucleic Acid Res. 1990; 18: 6339-6345.
31. Kopelowitz J, Hampe C, Goldman R, Reches M, Engelberg-Kulka H. Influence of codon context on UGA suppression and readthrough. J Mol Biol. 1992; 225: 261-269.
32. Gonzalez B, Ceciliani F, Galizzi A. Growth at low temperature suppresses read-through of the UGA stop codon during the expression of Bacillus subtillis flgM gene in Escherichia coli. J Biotechnol. 2003; 101: 173-180.