Factors that contribute to the immmunogenicity of therapeutic recombinant human proteins

Summary

Use of recombinant human proteins has revolutionized medicine by providing over 200 highly purified hormones and proteins that effectively treat many inherited and acquired peptide hormone and protein deficiencies. With the exception of therapeutic monoclonal antibodies, these biological medicines are synthesized by cultured cells using DNA sequences that would yield proteins with identical amino acid sequences as endogenous human proteins. Therefore, there was the broad expectation that recombinant human biological medicines would be non-immunogenic in patients capable of synthesizing even sub-optimal levels of these therapeutic proteins to which they are innately tolerant. However, the widespread clinical use of recombinant human proteins has demonstrated that nearly all of them are immunogenic. This observation suggests that factors additional to differences in amino acid sequences of endogenous and biotherapeutic proteins contribute to the immunogenicity of therapeutic proteins. The main aim of this review is to summarize some of the factors that are known to contribute to the immunogenicity of recombinant therapeutic proteins.

Keywords

Therapeutic proteins - immunogenicity - recombinant human proteins

• References

• 1 Crommelin DJ, Storm G, Verrijk R. et al. Shifting paradigms: biopharmaceuticals versus low molecular weight drugs. Int J Pharm 2003; 266: 3-16.
• 2 Walsh G. Biopharmaceutical benchmarks 2006. Nat Biotechnol 2006; 24: 769-776.
• 3 Shankar G, Pendley C, Stein K-E. A risk-based bioanalytic strategy for the assessment of antibody immune responses against biological drugs. Nat Biotechnol 2007; 25: 555-561.
• 4 Sharma B. Immunogenicity of therapeutic proteins. Part 1: Impact of product handling. Biotechnol Advan 2007; 25: 310-317.
• 5 Schellekens H. The immunogenicity of biopharmaceuticals. Neurology 2003; 61: 11-12.
• 6 Schernthaner G. Immunogenicity and allergenic potential of animal and human insulins. Diabetes Care 1993; 16: 155-165.
• 7 Takano K, Shizume K. Clinical experience with somatrem in Japan. Acta Paediatr Scand Suppl 1986; 325: 19-24.
• 8 Suh BK, Jorgensen EV, Root AW. Facilitation of the growth. J Pediatr Endocrinol Metab 1995; 8: 97-102.
• 9 Jacquemin MG, Saint-Remy JM. Factor VIII immunogenicity. Haemophilia 1998; 4: 552-557.
• 10 Peces R, de la Torre M, Alcazar R. et al. Antibodies against Recombinant Human Erythropoietin in a Patient with Erythropoietin-Resistant Anemia. N Engl J Med 1996; 335: 523-524.
• 11 European Medicines Agency.. Guideline on similar biological medicinal products containing biotechnology derived proteins as active substance: non-clinical and clinical issues. Online 2006. Available at http://www.emea.eu.int/pdfs/human/biosimilar/4283205en.pdf.
• 12 Koren E, Zuckerman LA, Mire-Sluis AR. Immune responses to therapeutic proteins in humans-clinical significance, assessment and prediction. Curr Pharm Biotechnol 2002; 3: 349-360.
• 13 Center for Biologics Evaluation and Review.. Product Approval Information – Licensing Action, Etanercept. U.S. Food & Drug Administration. Online 2003. Available at http://www.fda.gov/cder/foi/label/2003/etanimm060503LB.pdf.
• 14 Perini P, Facchinetti A, Bulian P. et al. Interferon-beta (INF-beta) antibodies in interferon-beta1a- and interferon- beta1b-treated multiple sclerosis patients. Prevalence, kinetics, cross-reactivity, and factors enhancing interferon-beta immunogenicity in vivo. Eur Cytokine Net 2001; 12: 56-61.
• 15 Gribben JG, Devereu S, Thomas NS. et al. Development of antibodies to unprotected glycosylation sites on recombinant human GM-CSF. Lancet 1990; 335: 434-437.
• 16 Antonelli G, Giannelli G, Currenti M. et al. Antibodies to interferon (IFN) in hepatitis C patients relapsing while continuing recombinant IFN-alpha2 therapy. Clin and Exper Immunol 1996; 104: 384-387.
• 17 Zang YC, Ynag D, Hong J. et al. Immunoregulation and blocking antibodies induced by interferon beta treatment in MS. Neurology 2000; 55: 397-404.
• 18 Dharnidharka VR, Takemoto C, Ewenstein BM. et al. Membranous glomerulonephritis and nephrosis post factor. Pediatr Nephrol 1998; 12: 654-657.
• 19 Hunley TE, Corzo D, Dudek M. et al. Nephrotic syndrome complicating a-glucosidase replacement. Pediatrics 2004; 114: e532-e535.
• 20 Figlin RA, Itri LM. Anti-interferon antibodies: A perspective. Sem Hematol 1988; 25: 9-15.
• 21 Ryff JC, Schellekens H. Immunogenicity of rDNAderived pharmaceuticals. Trends in Pharmacol Sci 2002; 23: 254-256.
• 22 Rosenschein U, Lenz R, Radnay J. et al. Streptokinase immunogenicity in thrombolytic therapy for acute myocardial infarction. Isr J Med Sci 1991; 27: 541-545.
• 23 Li J, Yang C, Xia Y. et al. Thrombocytopenia caused by the development of antibodies to hrombopoietin. Blood 2001; 98: 3241-3248.
• 24 Casadevall N. Antibodies against rHuEPO: native and recombinant. Nephrology Dialysis Transplant 2002; 17: 42-47.
• 25 Lim LC. Acquired red cell aplasia in association with the use of recombinant erythropoietin in chronic renal failure. Hematology 2005; 10: 255-259.
• 26 Casadevall N, Nataf J, Viron B. et al. Pure red-cell aplasia and antierythropoietin antibodies in patients treated with recombinant erythropoietin. N Engl J Med 2002; 346: 469-475.
• 27 Gershon SK, Luksenburg H, Cote TR. et al. Pure red-cell aplasia and recombinant erythropoietin. N Engl J Med 2002; 346: 1584-1586.
• 28 Ragnhammar P, Wadhwa M. Neutralizing antibodies to granulocytemacrophage colony stimulating factor (GM-CSF) in carcinoma patients following GMCSF combination therapy. Med Oncol 1996; 13: 161-166.
• 29 Prümmer O. Treatment-induced antibodies to interleukin- 2. Biotherapy 1997; 10: 15-24.
• 30 Hartung HP, Munschauer F, Schellekens H. Significance of neutralizing antibodies to interferon beta during treatment of multiple sclerosis: expert opinions based on the Proceedings of an International Consensus Conference. Eur J Neurol 1995; 12: 588-601.
• 31 Öberg K, Alm G, Norheim L. et al. Treatment of malignant carcinoid tumors with recombinant interferon alfa-2b: Development of neutralizing interferon antibodies and possible loss of antitumor activity. Natl Cancer Inst 1989; 81: 531-535.
• 32 Sorensen PS, Ross C, Clemmesen KM. et al. Clinical importance of neutralizing antibodies against interferon- beta in patients with relapsing–remitting multiple sclerosis. Lancet 2003; 362: 1184-1191.
• 33 Schellekens H. Immunogenicity of therapeutic proteins: clinical implications and future prospects. Clin Ther 2002; 24: 1720-1740.
• 34 Janeway C. et al. Immunobiology.. 6th New York: Garland Science Publishing; 2005
• 35 Hausl C, Ahmed RU, Schwartz HP. et al. Preventing restimulation of memory B cells in hemophilia A: a potential new strategy for the treatment of antibody-dependent immune disorders. Blood 2004; 104: 115-122.
• 36 Qian J, Collins M, Sharpe AH. et al. Prevention and treatment of factor VIII inhibitors in murine hemophilia A. Blood 2000; 95: 1324-1329.
• 37 Reipert BM, Sasgary M, Ahmad Ru. et al. Blockade of CD40/CD40 ligand interactions prevents induction of factor VIII inhibitors in hemophilic mice but does not induce lasting immune tolerance. Thromb Haemost 2001; 86: 1345-1352.
• 38 Rossi G, Sarkar J, Scandella D. Long-term induction of immune tolerance after blockade of CD40-CD40L interaction in a mouse model of hemophilia A. Blood 2001; 97: 2750-2756.
• 39 Rosenberg AS. Effects of protein aggregates: an immunologic perspective. AAPS J 2006; 8: 501-507.
• 40 Martin F, Oliver A, Kearney J. Marginal zone and B1 B cells unite in the early response against T-independent blood-borne particulate antigen. Immunity 2001; 14: 617-629.
• 41 McHeyzer-Williams M, McHeyzer-Williams L, Panus J. et al. Helper T-cell-regulated B-cell immunity. Microb Infection 2003; 5: 205-212.
• 42 Chackerian B, Lenz P, Lowy DR. et al. Determinants of autoantibody induction by conjugated papillomavirus virus-like particles. J Immunol 2002; 169: 6120-6126.
• 43 Vos Q, Lees A, Wu ZQ. et al. B-cell activation by T-cell-independent type 2 antigens as an integral part of the humoral immune response to pathogenic microorganisms. Immunol Rev 2000; 176: 154-170.
• 44 Dintzis R, Okajrima M, Middleton MH. et al. The immunogenicity of soluble haptenated polymers is determined by molecular mass and hapten valen. J Immunol 1989; 143: 1239-1244.
• 45 Wagner CL, Schantz A, Barnathan E. et al. Consequences of immunogenicity to the therapeutic monoclonal antibodies. ReoPro and Remicade. Dev Biol (Basel) 2003; 112: 37-53.
• 46 Antonelli G, Dianzani F. Development of antibodies to interferon beta in patients: technical and biological aspects. Eur Cytokine Net 1999; 10: 413-422.
• 47 Oldenburg J, El-Maarri O, Schwaab R. Inhibitor development in correlation to factor VIII genotypes. Haemophilia 2002; 8: 23-29.
• 48 O’Connell NM. Factor XI deficiency from molecular genetics to clinical management. Blood Coagul Fibrinolysis 2003; 14 (Suppl. 01) 559-564.
• 49 Lawler P, White B, Pye S. et al. Successful use of recombinant factor VIIa in a patient with inhibitor secondary to severe factor XI deficiency. Haemophilia 2002; 8: 1245-148.
• 50 Tarnatino M, Aledort L. On the treatment of hemorrhage in patients with hemophilia and associated inhibitors. Transfusion 2001; 41: 1628-1629.
• 51 Schwaab R, Brackmann HH, Meyer C. et al. Haemophilia A. mutation type determines risk of inhibitor formation. Thromb and Haemost 1995; 74: 1402-1406.
• 52 Kemball-Cook G, Tuddenham EGD, Wacey AI. The factor VIII Structure and Mutation Resource Site: HAMSTeRS version 4. Nucleic Acids Res 1998; 26: 216-219.
• 53 Oldenburg J, Schroder J, Schmitt C. et al. Small deletion/ insertion mutations within poly-A-runs of factor VIII gene mitigate the severe haemophilia A phenotype. Thromb Haemost 1998; 79: 452-453.
• 54 Young M, Inaba H, Hoyer LW. et al. Partial correction of a severe molecular defect in haemophilia A, because of errors during expression of the factor VIII gene. Am J Human Genet 1997; 60: 565-573.
• 55 Scharrer I, Bray GL, Neutzling O. Incidence of inhibitors in haemophilia A patients – a review of recent studies of recombinant and plasma-derived factor VIII concentrates. Haemophilia 1999; 5: 145-154.
• 56 Cox G. The role of genetics in inhibitor formation. Thromb Haemost 1999; 82: 500-504.
• 57 Aly AM, Aledort LM, Lee TD. et al. Histocompatibility antigen patterns in haemophilic patients with factor VIII antibodies. Br J Haematol 1990; 76: 238-241.
• 58 Frommel D, Allain JP, Saint-Paul E. et al. HLA antigens and factor VIII antibody in classic hemophilia. European study group of factor VIII antibody. Thromb Haemost 1981; 46: 687-689.
• 59 Lippert LE, Fisher LM, Schook LB. Relationship of major histocompatibility complex class II genes to inhibitor antibody formation in haemophilia A. Thromb Haemost 1990; 64: 564-568.
• 60 Mayr WR, Lechner K, Niessher H. et al. Inhibitor development in correlation to factor VIII genotypes. Haemophilia 1984; 51: 293.
• 61 Oldenburg J, Pickard JK, Schwang R. et al. HLA genotype of patients with severe haemophilia A due to intron 22 inversion with and without inhibitors to factor VIII. Thromb Haemost 1997; 77: 238-242.
• 62 Barbosa MDFS, Vielmetter J, Chu S. et al. Clinical link between MHC class II haplotype and interferon –beta (IFN-β) immunogenicity. Clin Immunol 2006; 1189: 42-50.
• 63 Ottesen JL, Nilsson P, Jami J. et al. The potential immunogenicity of human insulin and insulin analogues evaluated in a transgenic mouse model. Diabetologia 1994; 37: 1178-1185.
• 64 Fireman P, Fineberg SE, Galloway JA. Development of IgE antibodies to human (recombinant DNA), porcine, and bovine insulins in diabetic subjects. Diabetes Care 1982; 5: 119-125.
• 65 Fineberg SE, Galloway JA, Fineberg NS. et al. Effects of species of origin, purification levels, and formulation on insulin immunogenicity. Diabetes 1983; 32: 592-599.
• 66 Kontsek P, Liptakova H, Kontsekova E. Immunogenicity of interferon-alpha 2 in therapy: structural and physiological aspects. Acta Virologica 1999; 43: 63-70.
• 67 Melian EB, Plosker GL. Interferon alfacon-1: a review of its pharmacology and therapeutic efficacy in the treatment of chronic hepatitis C. Drugs 2001; 61: 1661-1691.
• 68 Du X, Tang JG. Effects of deleting A19 tyrosine from insulin. Biochem Mol Biol Int 1998; 44: 507-513.
• 69 Stewart TA, Hollingshead PG, Pitts SL. et al. Transgenic mice as a model to test the immunogenicity of proteins altered by site-specific mutagenesis. Mol Biol Med 1989; 6: 275-281.
• 70 Hwang WYK, Foote J. Immunogenicity of engineered antibodies. Methods 2005; 36: 3-10.
• 71 Piro LD, White CA, Grillo-Lopez AJ. et al. Extended Rituximab (anti-CD20 monoclonal antibody) therapy for relapsed or refractory low-grade follicular non-Hodgkin’s lymphoma. Ann Oncol 1999; 10: 655-661.
• 72 Buist MR, Kenemans P, van Kamp GJ. et al. Minor human antibody response to a mouse and chimeric monoclonal antibody after a single i.v. infusion in ovarian carcinoma patients: a comparison of five assays. Cancer Immunol Immunother 1995; 40: 24-30.
• 73 Tcheng JE, Kereiakes DJ, Lincoff AM. Abciximols readministration: results of RCoPro Readministration Registry. Circulation 2001; 104: 870-875.
• 74 Baert F, Noman M, Vermeire G. et al. Influence of immunogenicity on the long-term efficicy of infliximab in Crohn’s disease. N Engl J Med 2003; 348: 601-608.
• 75 Ewensteini BM, Hoots WK, Lusher JM. et al. Inhibition of CD40 ligand (CD154) in the treatment of factor ViII inhibitors. Haematologica 2000; 85: 35-39.
• 76 Arnold DM, Kelton JG. Current options for the treatment of idiopathic thrombocytopenic purpura. Semin Hematol 2007; 44 (Suppl. 45) 512-523.
• 77 Walsh G. Therapeutic insulins and their large scale manufacturer. Appl Microbiol Biotechnol 2005; 67: 151-159.
• 78 Chu L, Robinson DK. Industrial choices for protein production by large-scale cell culture. Curr Opin Biotechnol 2001; 12: 180-187.
• 79 Swartz JR. Advances in Escherichia coli production of therapeutic proteins. Current Opin Biotechnology 2001; 12: 195-201.
• 80 Lin Y, Yang X, Chevrier MC. et al. Relationships between factor VIII:Ag and factor VIII in recombinant and plasma-derived factor VIII concentrates. Haemophilia 2005; 10: 459-469.
• 81 Ettingshausen CE, Kreuz W. Recombinant vs plasma- derived products, especially those with intact vWF, regarding inhibitor dev elopement. Haemophilia 2006; 12 (Suppl. 06) 102-106.
• 82 Gouw SC, van de Bom JG, Auerswald G. et al. Recombinant versus plasma-derived products and the development of inhibitors in previously untreated patients with severe hemophilia A: CANAL cohort study. Blood 2007; 1089: 4693-4697.
• 83 Goudemand J. Inhibitor development in haemophilia A: the role of von Willebrand factor/factor VIII concentrates. Haemophilia 207 13 (Suppl. 05) 47-51.
• 84 Kisker CT, Eisberg A, Schwartz B. Mononine Study Group. Prophylaxis in factor IX deficiency products and patient variation. Haemophilia 2003; 9: 279-284.
• 85 Wang L, Zoppe M, Hackeng TM. et al. A factor IX-deficient mouse model for hemophilia B gene therapy. Proc Nat Acad Sci USA 1997; 94: 11564-11566.
• 86 Hortelano G, Wang L, Xu N. et al. Sustained and therapeutic delivery of factor IX in nude haemophilia B mice by encapsulated C2C12 myoblasts: concurrent tumourigenesis. Haemophilia 2001; 7: 207-214.
• 87 Wen J, Ofosu FA, Hortelano G. Encapsulated primary myoblasts deliver sustained and therapeutic levels of human factor IX to hemophilic mice. J Gene Med 2006; 8: 362-369.
• 88 Butler M. Animal Cell Culture and Technology. 2nd ed. Oxford.: Bios Scientific; 2004
• 89 Neusüss C, Demelbauer U, Pelzing M. Glycoform characterization of intact erythropoietin by capillary electrophoresis-electrospray-time of flight-mass spectrometry. Electrophoresis 2005; 26: 1442-1450.
• 90 Venke S, Gro N-L, Peter T. Sugar profiling proves that human serum erythropoietin differs from recombinant human erythropoietin. Blood 2001; 98: 3626-3634.
• 91 Lasne F, de Ceaurriz J. Recombinant erythropoietin in urine. Nature 2000; 405: 635.
• 92 Galili U, Shohet SB, Kobrin E. et al. Man, apes, and Old World monkeys differ from other mammals in the expression of alpha-galactosyl epitopes on nucleated cells. J Biol Chem 1988; 263: 17755-17762.
• 93 Restelli V, Butler M. Cell engineering. Springer 2002; 3: 61-92.
• 94 Brooks S. Appropriate Glycosylation of Recombinant Proteins for Human Use: Implications of Choice of Expression System. Mol Biotechnol 2004; 28: 241-256.
• 95 Narhi LO, Arakwa T, Aoki KH. et al. The effect of carbohydrate on the structure and stability of erythropoietin. J Biol Chem 1991; 266: 23022-23026.
• 96 Wasley LC, Timony G, Murtha P. et al. The importance of N-and O-linked oligosaccharides for the biosynthesis and in vitro and in vivo biologic activities of erythropoietin. Blood 1991; 77: 2624-2632.
• 97 Storring PL. Assaying glycoprotein hormones–the influence of glycosylation on immunoreactivity. Tren Biotechnol 1992; 10: 427-432.
• 98 Takeuchi M, Takasai S, Miyazai H. et al. Comparative study of the asparagine-linked sugar chains of human erythropoietins purified from urine and the culture medium of recombinant Chinese hamster ovary cells. J Biol Chem 1988; 263: 3657-3663.
• 99 Goto M, Akai K, Murakami A. et al. Production of recombinant human erythropoietin in mammalian cells: Host–cell dependency of the biological activity of the cloned glycoprotein. Nat Biotechnol 1988; 6: 67-71.
• 100 Kagawa Y, Takasaki S, Utsumi J. et al. Comparative study of the asparagine-linked sugar chains of natural human interferon-beta 1 and recombinant human interferon-beta 1 produced by three different mammalian cells. J Biol Chem 1988; 263: 17508-17515.
• 101 Goochee CF. Bioprocess factors affecting glycoprotein oligosaccharide structure. Dev Biol Stand 1992; 76: 95-104.
• 102 Sheeley DM, Merrill BM, Taylor LC. Characterization of monoclonal antibody glycosylation: comparison of expression systems and identification of terminal alpha-linked galactose. Anal Biochem 1997; 247: 102-110.
• 103 Jenkins N, Curling EM. Glycosylation of recombinant proteins: problems and prospects. Enzyme Microb Technol 1994; 16: 354-364.
• 104 Gawlitzek M, Valley U, Nimtz MM. et al. Characterization of changes in the glycosylation pattern of recombinant proteins from BHK-21 cells due to different culture conditions. J Biotechnol 1995; 42: 117-131.
• 105 Schweikart F, Jones R, Jaton JC. et al. Rapid structural characterisation of a murine monoclonal IgA alpha chain: heterogeneity in the oligosaccharide structures at a specific site in samples produced in different bioreactor systems. J Biotechnol 1999; 69: 191-201.
• 106 Curling EM, Hayter PM, Baines AJ. et al. Recombinant human interferon-gamma. Differences in glycosylation and proteolytic processing lead to heterogeneity in batch culture. Biochem J 1990; 272: 333-337.
• 107 Nyberg GB, Balcarcel RR, Follstad DD. et al. Metabolic effects on recombinant interferon-gamma glycosylation in continuous culture of Chinese hamster ovary cells. Biotechnol Bioeng 1999; 62: 336-347.
• 108 Kinstler O, Molineux G, Treuheit M. et al. Mono- N-terminal poly(ethylene glycol)-protein conjugates. Adv Drug Deliv Rev 2002; 54: 447-485.
• 109 Kompella UB, Lee VHL. Pharmacokinetics of peptide and protein drugs. [book auth.] Lee V.H.L.. Peptide and Protein Drug Delivery.. New York: Marcel Dekker; 1991: 391.
• 110 Rabkin R, Dahl DC. Renal uptake and disposal of proteins and peptides. Biological Barriers in Protein Delivery.. New York: Plenum Press; 1993: 299.
• 111 Harris JM, Martin NE, Modi M. Pegylation: a novel process for modifying pharmacokinetics. J Clin Pharmacol 2000; 40: 539-551.
• 112 Delgado C, Francis GE, Fisher D. The uses and properties of PEG-linked proteins. Crit Rev Therap Drug Carrier Sys 1992; 9: 249-304.
• 113 Kartre K. The conjugation of proteins with poly(ethylene glycol) and other polymers. Adv Drug Deliv Rev 1993; 10: 91-114.
• 114 Roberts MJ, Bentley MD, Harris JM. Chemistry for peptide and protein PEGylation. Adv Drug Deliv Rev 2002; 54: 459-476.
• 115 Veronese FM. Peptide and protein PEGylation: a review of problems and solutions. Biomaterials 2001; 22: 405-417.
• 116 Chapman AP. PEGylated antibodies and antibody fragments for improved therapy: a review. Adv Drug Deliv Rev 2002; 54: 531-545.
• 117 Savoca KV, Abuchowski A, Van Es T. et al. Preparation of a non-immunogenic arginase by the covalent attachment of polyethylene glycol. Biochim Biophys Acta 1979; 578: 47-53.
• 118 Hershfield MS, Chaffee S, Koro-Johnson L. et al. Use of site-directed mutagenesis to enhance the epitope- shielding effect of covalent modification of proteins with polyethylene glycol. Proc Nat Acad Sci USA 1991; 88: 7185-7189.
• 119 Bailon P, Palleroni A, Schaffer CA. et al. Rational design of a potent, long lasting form of interferon: a 40kDa branched poly-ethylene glycol-conjugated interferon alpha-2a for the treatment of hepatitis C. Bioconj Chem 2001; 12: 195-202.
• 120 Reddy KR, Wright TL, Pokros PJ. et al. Efficacy and safety of pegylated (40-kd) interferon a-2a compared with interferon alpha-2a in noncirrhotic patients with chronic hepatitis C. Hepatology 2001; 33: 433-438.
• 121 Chirino AJ, Mire-Sluis A. Characterizing biological products and assessing comparability following manufacturing changes. Nat Biotechnol 2004; 22: 1383-1391.
• 122 Chance RE, Root MA, Galloway JA. The immunogenicity of insulin preparations. Acta Endocrinol Suppl 1976; 205: 185-198.
• 123 Wilson RM, Douglas CA, Tattersall RB. et al. Immunogenicity of highly purified bovine insulin: a comparison with conventional insulins. Diabetologia 1985; 28: 667-670.
• 124 Stubbe P, Ranke MB, Bierich JR. Incidence of growth hormone antibodies and presumed growth-attenuating antibodies in relation to various pituitary growth hormone preparations.. Munich; Urban and Schwarzenberg: 1999: 92-99.
• 125 Wadhwa M, Skog AL, Bird C. et al. Immunogenicity of granulocyte-macrophage colony-stimulating factor (GM-CSF) products in patients undergoing combination therapy with GM-CSF. Clin Cancer Res 1999; 5: 1353-1361.
• 126 Antonelli G, Simeoni E, Currenti M. et al. Interferon antibodies in patients with infectious diseases. Biotherapy. 1997: 107-114.
• 127 Wang W. Instability, stabilization and formulation of liquid protein pharmaceuticals. Int J Pharmaceut 1999; 185: 129-188.
• 128 Physician Package Insert Neulasta. Physician Package Insert Neulasta® (Pegfilgrastim). 2003
• 129 Reyes N, Ruiz L, Aroche K. et al. Stability of Ala 125 recombinant human interleukin-2 in solution. J Phar Pharmacol 2005; 57: 31-37.
• 130 Leppert P, Moore WV. Role of aggregated human growth hormone (hGH) in development of antibodies to hGH. J Clin Endocrinol Metab 1980; 51: 691-697.
• 131 Sharma B. Immunogenicity of therapeutic proteins. Part 2: Impact of container closures. Biotechnol Adv 2007; 25: 318-324.
• 132 Frokjaer S, Otzen DE. Protein drug stability: a formulation challenge. Nat Rev Drug Discov 2005; 4: 298-306.
• 133 Kreilgaard L, Frokjaer S. Effects of additives on the stability of recombinant human factor XIII during freeze-drying and storage in the dried solid. Arch Biochem Biophs 1998; 360: 121-134.
• 134 Krielgaard L, Jones LS, Randolph TW. et al. Effect of tween 20 on freeze-thawing- and agitation-induced aggregation of recombinant human factor XIII. J Pharm Sci 1998; 87: 1597-1603.
• 135 Katakam M, Banga AK. Use of poloxamer polymers to stabilize recombinant human growth hormone against various processing stresses. Pharma Dev Technol 1997; 2: 143-149.
• 136 Katakam M, Bell LN, Banga AK. Effect of surfactants on the physical stability of recombinant human growth hormone. J Pharm Sci 1995; 84: 713-716.
• 137 Bam NB, Cleland JL, Yang J. et al. Tween protects recombinant human growth hormone against agitation-induced damage via hydrophobic interactions. J Pharma Sci 1998; 87: 1554-1559.
• 138 Brange J, Anderson L, Laursen ED. et al. Toward understanding insulin fibrillation. J Pharm Sci 1997; 86: 517-525.
• 139 Kwon YM, Boudys M, Knutson K. et al. In situ study of insulin aggregation induced by water-organic solvent interface. Pharm Res 2001; 18: 1754-1759.
• 140 Bam NB, Randolph TW, Cleland JL. Stability of protein formulations: investigation of surfactant effects by a novel EPR spectroscopic technique. Pharm Res 1995; 12: 2-11.
• 141 Webb SD, Cleland JL, Carpenter JF. A new mechanism for decreasing aggregation of recombinant human interferon-gamma by a surfactant: slowed dissolution of lyophilized formulations in a solution containing 0.03% polysorbate 20. J Pharm Sci 002 91: 543-558.
• 142 Webb SD, Golledge SL, Cleland JL. et al. Surface adsorption of recombinant human interferon-gamma in lyophilized and spray-lyophilized formulations. J Pharm Sci 2002; 91: 1474-1487.
• 143 Maa YF, Hsu CC. Protein denaturation by combined effect of shear and air–liquid interface. Biotechnol Bioeng 1997; 54: 503-512.
• 144 Treuheit MJ, Kosky AA, Brems DN. Inverse relationship of protein concentration and aggregation. Pharm Res 2002; 19: 511-516.
• 145 Sadhale Y, Shah JC. Stabilization of insulin against agitation-induced aggregation by the GMO cubic phase gel. Int J Pharm 1999; 191: 51-64.
• 146 Hochuli E. Interferon immunogenicity: technical evaluation of interferon-alpha 2a. J Interferon Cytokine Res 1997; 17: S15-21.
• 147 Emily HA, Wei W, John W. Peroxide formation in polysorbate 80 and protein stability. J Pharm Sci 2002; 91: 2252-2264.
• 148 Lam XM, Yang JY, Cleland JL. Antioxidants for prevention of methionine oxidation in recombinant monoclonal antibody HER2. J Pharm Sci 1997; 86: 1250-1255.
• 149 Johnston TP. Adsorption of recombinant human granulocyte colony stimulating factor (rhG-CSF) to polyvinyl chloride, polypropylene, and glass: effect of solvent additives. PDA J Pharm Sci Tech 1996; 50: 238-245.
• 150 Jones LS, Kaufmann A, Middaugh CR. Silicone oil induced aggregation of proteins. J Pharm Sci 2005; 94: 918-927.
• 151 Mizutani T. Decreased activity of proteins absorbed onto glass surfaces with porous glass as a reference. J Pharm Sci 1980; 69: 279-282.
• 152 Sharma B. Immunogenicity of therapeutic proteins. Part 3: Impact of manufacturing changes. Biotechnol Adv 2007; 25: 325-331.
• 153 Sharma B. Technical investigations into the cause of the increased incidence of antibody-mediated pure red cell aplasia associated with Eprex®. Eur J Hosp Pharm 2004; 5: 86-91.
• 154 Cleland JL, Powell MF, Shire SJ. The development of stable protein formulations: a close look at protein aggregation, deamidation, and oxidation. Crit Rev Drug Carrier Sys 1993; 10: 307-377.
• 155 Shihong L, Christian S, Borchardt R. Chemical instability of protein pharmaceuticals: Mechanisms of oxidation and strategies for stabilization. Biotechnol and Bioeng 1995; 48: 490-500.
• 156 Hermeling S, Crommelin DJ, Schellekens H. et al. Structure-immunogenicity relationships of therapeutic proteins. Pharma Res 2004; 21: 897-903.
• 157 Ross C, Clemmensen KM, Svenson M. et al. Immunogenicity of interferon-beta in multiple sclerosis patients: Influence of preparation, dosage, dose frequency, and route of administration. Ann Neurol 2000; 48: 706-712.
• 158 Fineberg SE, Kawabata T. Finio-Kent et al. Antibody response to inhaled insulin in patients with type 1 or type 2 diabetes. An analysis of initial phase II and II inhaled insulin (Exubera) trials and a two-year extension. J Clin Endocrinol Metab 2005; 90: 3287-3294.