Fusion Protein Linkers: Effects on Production, Bioactivity, and Pharmacokinetics

Fusion Protein Technologies for Biopharmaceuticals: Applications and Challenges

Volumn , Issue , 2013, Pages 57-73

  Chen, Xiaoying ^a   Zaro, Jennica ^b   Shen, Wei Chiang ^b  

^a PFIZER INC   (United States)

^b UNIVERSITY OF SOUTHERN CALIFORNIA   (United States)

Author keywords

Empirical linkers; FP linkers; Linker functionality; Linkers, PK of FPs; Production bioactivity

Indexed keywords

EID: 84886143139     PISSN: None     EISSN: None     Source Type: Book    
DOI: 10.1002/9781118354599.ch4     Document Type: Chapter

References (73)

1. Yuste R. (2005) Fluorescence microscopy today. Nat. Meth. 2, 902-904.
2. Terpe K. (2003) Overview of tag protein fusions: from molecular and biochemical fundamentals to commercial systems. Appl. Microbiol. Biotechnol. 60, 523-533.
3. Lu P, Feng MG. (2008) Bifunctional enhancement of a betaglucanase- xylanase fusion enzyme by optimization of peptide linkers. Appl. Microbiol. Biotechnol. 79, 579-587.
4. Schmidt SR. (2009) Fusion-proteins as biopharmaceuticals- applications and challenges. Curr. Opin. Drug Discov. Dev. 12, 284-295.
5. Kreitman RJ. (2006) Immunotoxins for targeted cancer therapy. AAPS J. 8, E532-E551.
6. Pardridge WM. (2010) Biopharmaceutical drug targeting to the brain. J. Drug Target. 18, 157-167.
7. Kim B, Zhou J, Martin B, Carlson O, Maudsley S, Greig N, et al. (2010) Transferrin fusion technology: a novel approach to prolong biological half-life of insulinotropic peptides. J. Pharmacol. Exp. Ther. Epub (DOI: 10.1124/jpet.110.166470).
8. Weimer T,WormsbacherW, Kronthaler U, LangW, Liebing U, Schulte S. (2008) Prolonged in-vivo half-life of factor VIIa by fusion to albumin. Thromb. Haemost. 99, 659-667.
9. Dumont JA, Low SC, Peters RT, Bitonti AJ. (2006) Monomeric Fc fusions: impact on pharmacokinetic and biological activity of protein therapeutics. BioDrugs 20, 151-160.
10. Schellenberger V,Wang C, Geething N, Spink B, Campbell A, To W, et al. (2009) A recombinant polypeptide extends the in vivo half-life of peptides and proteins in a tunable manner. Nat. Biotechnol. 27, 1186-1190.
11. Zhao H, Yao X, Xue C, Wang Y, Xiong X, Liu Z. (2008) Increasing the homogeneity, stability and activity of human serum albumin and interferon-alpha2b fusion protein by linker engineering. Protein Expr. Purif. 61, 73-77.
12. Amet N, Lee HF, Shen WC. (2009) Insertion of the designed helical linker led to increased expression of tf-based fusion proteins. Pharm. Res. 26, 523-528.
13. Bai Y, Ann DK, Shen WC. (2005) Recombinant granulocyte colony-stimulating factor-transferrin fusion protein as an oral myelopoietic agent. Proc. Natl. Acad. Sci. USA 102, 7292-7296.
14. Bai Y, Shen WC. (2006) Improving the oral efficacy of recombinant granulocyte colony-stimulating factor and transferrin fusion protein by spacer optimization. Pharm. Res. 23, 2116-2121.
15. Robinson CR, Sauer RT. (1998) Optimizing the stability of single-chain proteins by linker length and composition mutagenesis. Proc. Natl. Acad. Sci. USA 95, 5929-5934.
16. Wriggers W, Chakravarty S, Jennings PA. (2005) Control of protein functional dynamics by peptide linkers. Biopolymers 80, 736-746.
17. George R, Heringa J. (2002) An analysis of protein domain linkers: their classification and role in protein folding. Protein Eng. 15, 871-879.
18. Argos P. (1990) An investigation of oligopeptides linking domains in protein tertiary structures and possible candidates for general gene fusion. J. Mol. Biol. 211, 943-958.
19. Eisenberg D. (1984) Three-dimensional structure of membrane and surface proteins. Annu. Rev. Biochem. 53, 595-623.
20. Aurora R, Creamer T, Srinivasan R, Rose G. (1997) Local interactions in protein folding: lessons from the alpha-helix. J. Biol. Chem. 272, 1413-1416.
21. Williamson MP. (1994) The structure and function of prolinerich regions in proteins. Biochem. J. 297(Pt 2), 249-260.
22. Hagemeyer CE, von Zur Muhlen C, von Elverfeldt D, Peter K. (2009) Single-chain antibodies as diagnostic tools and therapeutic agents. Thromb. Haemost. 101, 1012-1019.
23. Huston J, Levinson D, Mudgett-Hunter M, Tai M, Novotny J, Margolies M, et al. (1988) Protein engineering of antibody binding sites: recovery of specific activity in an anti-digoxin single-chain Fv analogue produced in Escherichia coli. Proc. Natl. Acad. Sci. USA 85, 5879-5883.
24. Holliger P, Prospero T, Winter G. (1993) "Diabodies": small bivalent and bispecific antibody fragments. Proc. Natl. Acad. Sci. USA 90, 6444-6448.
25. Iliades P, Kortt A, Hudson P. (1997) Triabodies: single chain Fv fragments without a linker form trivalent trimers. FEBS Lett. 409, 437-441.
26. Amet N,WangW, Shen WC. (2010) Human growth hormonetransferrin fusion protein for oral delivery in hypophysectomized rats. J. Control. Release 141, 177-182.
27. Xia CQ, Wang J, Shen WC. (2000) Hypoglycemic effect of insulin-transferrin conjugate in streptozotocin-induced diabetic rats. J. Pharmacol. Exp. Ther. 295, 594-600.
28. Hu W, Li F, Yang X, Li Z, Xia H, Li G, et al. (2004) A flexible peptide linker enhances the immunoreactivity of two copies HBsAg preS1 (21-47) fusion protein. J. Biotechnol. 107, 83-90.
29. Bergeron LM, Gomez L, Whitehead TA, Clark DS. (2009) Self-renaturing enzymes: design of an enzyme-chaperone chimera as a new approach to enzyme stabilization. Biotechnol. Bioeng. 102, 1316-1322.
30. Bird R, Hardman K, Jacobson J, Johnson S, Kaufman B, Lee S, et al. (1988) Single-chain antigen-binding proteins. Science 242, 423-426.
31. Sabourin M, Tuzon C, Fisher T, Zakian V. (2007) A flexible protein linker improves the function of epitope-tagged proteins in Saccharomyces cerevisiae. Yeast 24, 39-45.
32. Waldo G, Standish B, Berendzen J, Terwilliger T. (1999) Rapid protein-folding assay using green fluorescent protein. Nat. Biotechnol. 17, 691-695.
33. Maeda Y, Ueda H, Kazami J, Kawano G, Suzuki E, Nagamune T. (1997) Engineering of functional chimeric protein GVargula luciferase. Anal. Biochem. 249, 147-152.
34. Arai R, Ueda H, Kitayama A, Kamiya N, Nagamune T. (2001) Design of the linkers which effectively separate domains of a bifunctional fusion protein. Protein Eng. 14, 529-532.
35. Arai R, Wriggers W, Nishikawa Y, Nagamune T, Fujisawa T. (2004) Conformations of variably linked chimeric proteins evaluated by synchrotron X-ray small-angle scattering. Proteins 57, 829-838.
36. Marqusee S, Baldwin RL. (1987) Helix stabilization by Glu-Lys+ salt bridges in short peptides of de novo design. Proc. Natl. Acad. Sci. USA 84, 8898-8902.
37. Werner S, Marillonnet S, Hause G, Klimyuk V, Gleba Y. (2006) Immunoabsorbent nanoparticles based on a tobamovirus displaying protein A. Proc. Natl. Acad. Sci. USA 103, 17678-17683.
38. Takamatsu N,Watanabe Y, Yanagi H, Meshi T, Shiba T, Okada Y. (1990) Production of enkephalin in tobacco protoplasts using tobacco mosaic virus RNA vector. FEBS Lett. 269, 73-76.
39. Turner S, Russell G, Williamson M, Guest J. (1993) Restructuring an interdomain linker in the dihydrolipoamide acetyltransferase component of the pyruvate dehydrogenase complex of Escherichia coli. Protein Eng. 6, 101-108.
40. Erni B. (1989) Glucose transport in Escherichia coli. FEMS Microbiol. Rev. 5, 13-23.
41. Bhandari D, Levine B, Trayer I, Yeadon M. (1986) 1H-NMR study of mobility and conformational constraints within the proline-rich N-terminal of the LC1 alkali light chain of skeletal myosin. Correlation with similar segments in other protein systems. Eur. J. Biochem. 160, 349-356.
42. Evans J, Levine B, Trayer I, Dorman C, Higgins C. (1986) Sequence-imposed structural constraints in the TonB protein of E. coli. FEBS Lett. 208, 211-216.
43. Morris A, MacArthur M, Hutchinson E, Thornton J. (1992) Stereochemical quality of protein structure coordinates. Proteins 12, 345-364.
44. McCormick A, Thomas M, Heath A. (2001) Immunization with an interferon-gamma-gp120 fusion protein induces enhanced immune responses to human immunodeficiency virus gp120. J. Infect. Dis. 184, 1423-1430.
45. Chen X, Bai Y, Zaro J, Shen WC. (2010) Design of an in vivo cleavable disulfide linker in recombinant fusion proteins. Biotechniques 49, 513-518.
46. Schulte S. (2009) Half-life extension through albumin fusion technologies. Thromb. Res. 124Suppl. 2, S6-8.
47. Vessillier S, Adams G, Chernajovsky Y. (2004) Latent cytokines: development of novel cleavage sites and kinetic analysis of their differential sensitivity to MMP-1 and MMP-3. Protein Eng. Des. Sel. 17, 829-835.
48. Wakefield LM, Winokur TS, Hollands RS, Christopherson K, Levinson AD, Sporn MB. (1990) Recombinant latent transforming growth factor beta 1 has a longer plasma half-life in rats than active transforming growth factor beta 1, and a different tissue distribution. J. Clin. Invest. 86, 1976-1984.
49. Martel-Pelletier J, Welsch DJ, Pelletier JP. (2001) Metalloproteases and inhibitors in arthritic diseases. Best Pract. Res. Clin. Rheumatol. 15, 805-829.
50. Lampert K, Machein U, Machein MR, Conca W, Peter HH, Volk B. (1998) Expression of matrix metalloproteinases and their tissue inhibitors in human brain tumors. Am. J. Pathol. 153, 429-437.
51. Yong VW, Krekoski CA, Forsyth PA, Bell R, Edwards DR. (1998) Matrix metalloproteinases and diseases of the CNS. Trends Neurosci. 21, 75-80.
52. Leppert D, Lindberg RL, Kappos L, Leib SL. (2001) Matrix metalloproteinases: multifunctional effectors of inflammation in multiple sclerosis and bacterial meningitis. Brain Res. Rev. 36, 249-257.
53. Pepinsky RB, LePage DJ, Gill A, Chakraborty A, Vaidyanathan S, Green M, et al. (2001) Improved pharmacokinetic properties of a polyethylene glycol-modified form of interferon- beta-1a with preserved in vitro bioactivity. J. Pharm. Exp. Ther. 297, 1059-1066.
54. Peng KW, Morling FJ, Cosset FL, Murphy G, Russell SJ. (1997) A gene delivery system activatable by diseaseassociated matrix metalloproteinases. Hum. Gene Ther. 8, 729-738.
55. Nagase H, Fields GB. (1996) Human matrix metalloproteinase specificity studies using collagen sequence-based synthetic peptides. Biopolymers 40, 399-416.
56. Liu S, Netzel-Arnett S, Birkedal-Hansen H, Leppla SH. (2000) Tumor cell-selective cytotoxicity of matrix metalloproteinaseactivated anthrax toxin. Cancer Res. 60, 6061-6067.
57. Thomas G. (2002) Furin at the cutting edge: from protein traffic to embryogenesis and disease. Nat. Rev. Mol. Cell Biol. 3, 753-766.
58. Goyal A, Batra JK. (2000) Inclusion of a furin-sensitive spacer enhances the cytotoxicity of ribotoxin restrictocin containing recombinant single-chain immunotoxins. Biochem. J. 345Pt 2, 247-254.
59. Wang T, Zhao J, Ren JL, Zhang L, Wen WH, Zhang R, et al. (2007) Recombinant immunoproapoptotic proteins with furin site can translocate and kill HER2-positive cancer cells. Cancer Res. 67, 11830-11839.
60. Widera A, Norouziyan F, Shen WC. (2003) Mechanisms of TfR-mediated transcytosis and sorting in epithelial cells and applications toward drug delivery. Adv. Drug Deliv. Rev. 55, 1439-1466.
61. Shah D, Shen WC. (1994) The establishment of polarity and enhanced transcytosis of transferrin receptors in enterocytelike Caco-2 cells. J. Drug Target. 2, 93-99.
62. Duttaroy A, Kanakaraj P, Osborn B, Schneider H, Pickeral O, Chen C, et al. (2005) Development of a long-acting insulin analog using albumin fusion technology. Diabetes 54, 251-258.
63. Halpern W, Riccobene T, Agostini H, Baker K, Stolow D, Gu M, et al. (2002) Albugranin, a recombinant human granulocyte colony stimulating factor (G-CSF) genetically fused to recombinant human albumin induces prolonged myelopoietic effects in mice and monkeys. Pharm. Res. 19, 1720-1729.
64. Zhao H, Xue C, Wang Y, Li X, Xiong X, Yao X, et al. (2007) Circumventing the heterogeneity and instability of human serum albumin-interferon-alpha2b fusion protein by altering its orientation. J. Biotechnol. 131, 245-252.
65. Leader B, Baca Q, Golan D. (2008) Protein therapeutics: a summary and pharmacological classification. Nat. Rev. Drug Discov. 7, 21-39.
66. Osborn B, Olsen H, Nardelli B,Murray J, Zhou J, Garcia A, et al. (2002) Pharmacokinetic and pharmacodynamic studies of a human serum albumin-interferon-alpha fusion protein in cynomolgus monkeys. J. Pharmacol. Exp. Ther. 303, 540-548.
67. Muller N, Schneider B, Pfizenmaier K, Wajant H. (2010) Superior serum half life of albumin tagged TNF ligands. Biochem. Biophys. Res. Commun. 396, 793-799.
68. Zhu D, Kepley CL, Zhang M, Zhang K, Saxon A. (2002) A novel human immunoglobulin Fc gamma Fc epsilon bifunctional fusion protein inhibits Fc epsilon RI-mediated degranulation. Nat. Med. 8, 518-521.
69. Manoukian G, Hagemeister F. (2009) Denileukin diftitox: a novel immunotoxin. Exp. Opin. Biol. Ther. 9, 1445-1451.
70. van Kerkhof P, Strous G. (2001) The ubiquitin-proteasome pathway regulates lysosomal degradation of the growth hormone receptor and its ligand. Biochem. Soc. Trans. 29, 488-493.
71. Grant B, Donaldson J. (2009) Pathways and mechanisms of endocytic recycling. Nat. Rev. Mol. Cell Biol. 10, 597-608.
72. Muller-Eberhard U, Liem H, Grasso J, Giffhorn-Katz S, DeFalco M, Katz N. (1988) Increase in surface expression of transferrin receptors on cultured hepatocytes of adult rats in response to iron deficiency. J. Biol. Chem. 263, 14753-14756.
73. Crasto CJ, Feng JA. (2000) LINKER: a program to generate linker sequences for fusion proteins. Protein Eng. 13, 309-312.