Insights on the Formulation of Recombinant Proteins

  • Rita Ribeiro
  • Teresa Raquel Abreu
  • Ana Catarina Silva
  • João Gonçalves
  • João Nuno MoreiraEmail author
Part of the Advances in Biochemical Engineering/Biotechnology book series (ABE, volume 171)


Recombinant proteins are large and complex molecules, whose therapeutic activity highly depends on their structure. Formulation of biopharmaceuticals aims at stabilizing protein conformation, promoting its efficacy, and preventing safety concerns, such as immunogenicity. Currently, the rational design of formulations is possible due to the availability of several techniques for molecule characterization and an array of both well-known and new excipients. Also, high-throughput technologies and Quality by Design approaches are trending and have been contributing to the advancement of the field. Still, there is a search for alternatives that ensure quality of the medicines through its life cycle, particularly for highly concentrated formulations, such as monoclonal antibodies. There is also a demand for strategies that improve protein delivery and more comfortable administration to the patients, especially with the arising of recombinant proteins in the treatment of chronic diseases, such as autoimmune conditions or heart diseases. In this chapter, current and future advancements regarding recombinant protein formulation and its impact in drug development and approval will be addressed.

Graphical Abstract


Formulation Immunogenicity Recombinant proteins Stability 



Rita Ribeiro is a student of the Ph.D. Program in Pharmaceutical Sciences from the Faculty of Pharmacy, University of Coimbra, and a recipient of the fellowship SFRH/BD/121935/2016 from the Portuguese Foundation for Science and Technology. This work was funded by Portuguese National funds via FCT – Fundação para a Ciência e a Tecnologia, I.P. – under projects Cancel Stem (reference POCI-01-0145-FEDER-016390), CENTRO-01-0145-FEDER-000012 (HealthyAging2020), Euronanomed2 (FCT reference ENMed/0005/2015), and CNC.IBILI (FCT reference UID/NEU/04539/2019).


  1. 1.
    Dill KA, Maccallum JL (2012) The protein-folding problem, 50 years on. Science 338:1042–1047PubMedGoogle Scholar
  2. 2.
    Dobson CM (2003) Protein folding and misfolding. Nature 426:884–890PubMedGoogle Scholar
  3. 3.
    Goswami S, Wang W, Arakawa T, Ohtake S (2013) Developments and challenges for mAb-based therapeutics. Antibodies 89:452–500Google Scholar
  4. 4.
    Krause ME, Sahin E (2019) Chemical and physical instabilities in manufacturing and storage of therapeutic proteins. Curr Opin Biotechnol 60:159–167PubMedGoogle Scholar
  5. 5.
    Randolph TW, Carpenter JF (2007) Engineering challenges of protein formulations. Am Inst Chem Eng J 53:1902–1907Google Scholar
  6. 6.
    Pisal DS, Kosloski MP, Balu-iyer SV (2009) Delivery of therapeutic proteins. J Pharm Sci 99:2557–2575Google Scholar
  7. 7.
    Manning MC, Chou DK, Murphy BM, Payne RW, Katayama DS (2010) Stability of protein pharmaceuticals: an update. Pharm Res 27:544–575Google Scholar
  8. 8.
    Soulby AJ, Heal JW, Barrow MP, Roemer RA, Connor PBO (2015) Does deamidation cause protein unfolding? A top-down tandem mass spectrometry study. Protein Sci 24:850–860PubMedPubMedCentralGoogle Scholar
  9. 9.
    Tyler-cross R, Schirchs V (1991) Effects of amino acid sequence, buffers, and ionic strength on the rate and mechanism of deamidation of asparagine residues in small peptides. J Biol Chem 266:22549–22556PubMedGoogle Scholar
  10. 10.
    Diepold K et al (2012) Simultaneous assessment of asp isomerization and asn deamidation in recombinant antibodies by LC-MS following incubation at elevated temperatures. PLoS One 7:1–11Google Scholar
  11. 11.
    Gervais D (2016) Protein deamidation in biopharmaceutical manufacture: understanding, control and impact. J Chem Technol Biotechnol 91:569–575Google Scholar
  12. 12.
    Parkins DA, Lashmar UT (2000) The formulation of biopharmaceutical products. Pharm Sci Technol Today 3:129–137PubMedGoogle Scholar
  13. 13.
    Stadtman ER (1990) Metal ion-catalyzed oxidation of proteins: biochemical mechanism and biological consequences. Free Radic Biol Med 9:315–325PubMedGoogle Scholar
  14. 14.
    Ha E, Wang WEI, Wang YJ (2002) Peroxide formation in polysorbate 80 and protein stability. J Pharm Sci 91:2252–2264PubMedGoogle Scholar
  15. 15.
    Torosantucci R, Schöneich C, Jiskoot W (2014) Oxidation of therapeutic proteins and peptides: structural and biological consequences. Pharm Res 31:541–553PubMedGoogle Scholar
  16. 16.
    Li S, Schoneich C, Borchardt RT (1995) Chemical instability of protein pharmaceuticals: mechanisms of oxidation and strategies for stabilization. Biotechnol Bioeng 48:490–500PubMedGoogle Scholar
  17. 17.
    Wang WEI et al (2006) Antibody structure, instability, and formulation. J Pharm Sci 96:1–26Google Scholar
  18. 18.
    Kiese S, Papppenberger A, Friess W, Mahler H (2008) Shaken, not stirred: mechanical stress testing of an IgG1 antibody. J Pharm Sci 97:4347–4366PubMedGoogle Scholar
  19. 19.
    Wang W (1999) Instability, stabilization, and formulation of liquid protein pharmaceuticals. Int J Pharm 185:129–188CrossRefGoogle Scholar
  20. 20.
    Carpenter BJF, Kendrick BS, Chang BS, Manning MC, Randolph TW (1999) Inhibition of stressed-induced aggregation of protein therapeutics. Methods Enzymol 309:236–255PubMedGoogle Scholar
  21. 21.
    Kueltzo LA, Wang WEI, Randolph TW, Carpenter JF (2008) Effects of solution conditions, processing parameters, and container materials on aggregation of a monoclonal antibody during freeze–thawing. J Pharm Sci 97:1801–1812PubMedGoogle Scholar
  22. 22.
    Dias CL et al (2010) The hydrophobic effect and its role in cold denaturation. Cryobiology 60:91–99PubMedGoogle Scholar
  23. 23.
    Chi EY, Krishnan S, Randolph TW, Carpenter JF (2003) Physical stability of proteins in aqueous solution: mechanism and driving forces in nonnative protein aggregation. Pharm Res 20:1325–1336PubMedGoogle Scholar
  24. 24.
    Joubert MK et al (2012) Highly aggregated antibody therapeutics can enhance the in vitro innate and late-stage T-cell immune responses. J Biol Chem 287:25266–25279PubMedPubMedCentralGoogle Scholar
  25. 25.
    Andersen CB, Manno M, Rischel C, Thórólfsson M, Martorana V (2010) Aggregation of a multidomain protein: a coagulation mechanism governs aggregation of a model IgG1 antibody under weak thermal stress. Protein Sci 19:279–290PubMedGoogle Scholar
  26. 26.
    Frokjaer S, Otzen DE (2005) Protein drug stability: a formulation challenge. Nat Rev Drug Discov 4:298–306PubMedGoogle Scholar
  27. 27.
    Chiti F, Stefani M, Ramponi G, Dobson CM (2003) Rationalization of the effects of mutations on peptide and protein aggregation rates. Nature 424:805–808PubMedGoogle Scholar
  28. 28.
    Nielsen L, Frokjaer S, Brange J, Uversky VN, Fink AL (2001) Probing the mechanism of insulin fibril formation with insulin mutants. Biochemistry 40:8397–8409PubMedGoogle Scholar
  29. 29.
    Agrawal NJ et al (2011) Aggregation in protein-based biotherapeutics: computational studies and tools to identify aggregation-prone regions. J Pharm 100:5081–5095Google Scholar
  30. 30.
    Obrezanova O et al (2015) Aggregation risk prediction for antibodies and its application to biotherapeutic development. MAbs 7:352–363PubMedPubMedCentralGoogle Scholar
  31. 31.
    Pandya A, Howard MJ, Zloh M, Dalby PA (2018) An evaluation of the potential of NMR spectroscopy and computational modelling methods to inform biopharmaceutical formulations. Pharmaceutics 10:1–24Google Scholar
  32. 32.
    ICH (1999) ICH Q6B – specifications: test procedures and acceptance criteria for biotechnological/biological products. 1–16Google Scholar
  33. 33.
    Beck A, Wagner-rousset E, Ayoub D, van Dorsselaer A, Sanglier-cianférani S (2013) Characterization of therapeutic antibodies and related products. Anal Chem 85:715–736PubMedGoogle Scholar
  34. 34.
    Crommelin D (2013) Formulation of biotech products, including biopharmaceutical considerations. In: Pharmaceutical biotechnology. CRC Press, Boca Raton, pp 69–96Google Scholar
  35. 35.
    Angkawinitwong U, Sharma G, Khaw PT, Brocchini S (2015) Solid-state protein formulations. Ther Deliv 6:59–82PubMedGoogle Scholar
  36. 36.
    Solá RJ, Griebenow KAI (2010) Effects of glycosylation on the stability of protein pharmaceuticals. J Pharm Sci 98:1223–1245Google Scholar
  37. 37.
    Jorgensen L, Hostrup S, Moeller EH, Grohganz H (2009) Recent trends in stabilising peptides and proteins in pharmaceutical formulation – considerations in the choice of excipients. Expert Opin Drug Deliv 6:1219–1230PubMedGoogle Scholar
  38. 38.
    Peters B et al (2016) Effects of cooling rate in microscale and pilot scale freeze-drying – variations in excipient polymorphs and protein secondary structure. Eur J Pharm Sci 95:72–81PubMedGoogle Scholar
  39. 39.
    Gervasi V et al (2018) Parenteral protein formulations: an overview of approved products within the European Union. Eur J Pharm Biopharm 131:8–24PubMedGoogle Scholar
  40. 40.
    Garidel P, Kuhn AB, Schäfer LV, Karow-zwick AR, Blech M (2017) High-concentration protein formulations: how high is high. Eur J Pharm Biopharm 119:353–360PubMedGoogle Scholar
  41. 41.
    Hawe A, Frieß W (2007) Formulation development for hydrophobic therapeutic proteins. Pharm Dev Technol 12:223–237PubMedGoogle Scholar
  42. 42.
    Tedeschi G, Mangiagalli M, Chmielewska S, Natalello A, Brocca S (2017) Aggregation properties of a disordered protein are tunable by pH and depend on its net charge per residue. Biochim Biophys Acta 1861:2543–2550Google Scholar
  43. 43.
    Hopkins E, Sharma S (2019) Physiology, acid base balance. StatPearls. Accessed 12 Oct 2019
  44. 44.
    Roethlisberger D, Mahler H, Altenburger U, Pappenberger A (2016) If euhydric and isotonic do not work, what are acceptable pH and osmolality for parenteral drug dosage forms? J Pharm Sci 106:1–11Google Scholar
  45. 45.
    Bahrenburg S, Karow AR, Garidel P (2015) Buffer-free therapeutic antibody preparations provide a viable alternative to conventionally buffered solutions: from protein buffer capacity prediction to bioprocess applications. Biotechnol J 10:610–622PubMedGoogle Scholar
  46. 46.
    Shire SJ (2009) Formulation and manufacturability of biologics. Curr Opin Biotechnol 20:708–714PubMedGoogle Scholar
  47. 47.
    Arakawa T, Tsumoto K, Kita Y, Chang B, Ejima D (2007) Biotechnology applications of amino acids in protein purification and formulations. Amino Acids 33:587–605PubMedGoogle Scholar
  48. 48.
    Shukla D, Trout BL (2011) Understanding the synergistic effect of arginine and glutamic acid mixtures on protein solubility. J Phys Chem 115:11831–11839Google Scholar
  49. 49.
    Al-hussein A, Gieseler H (2013) Investigation of histidine stabilizing effects on LDH during freeze-drying. J Pharm Sci 102:813–826PubMedGoogle Scholar
  50. 50.
    Wang W (2000) Lyophilization and development of solid protein pharmaceuticals. Int J Pharm 203:1–60PubMedGoogle Scholar
  51. 51.
    Pikal MJ, Dellerman KM, Roy ML, Riggin RM (1991) The effects of formulation variables on the stability of freeze-dried human growth hormone. Pharm Res 8:427–436PubMedGoogle Scholar
  52. 52.
    Akers MJ (2002) Excipient-drug interactions in parenteral formulations. J Pharm Sci 91:2283–2300PubMedGoogle Scholar
  53. 53.
    Khan TA, Mahler H, Kishore RSK (2015) Key interactions of surfactants in therapeutic protein formulations: a review. Eur J Pharm Biopharm 97:60–67PubMedGoogle Scholar
  54. 54.
    Goyal MK, Roy I, Amin A, Banerjee UC, Bansal AK (2010) Stabilization of lysozyme by benzyl alcohol: surface tension and thermodynamic parameters. J Pharm Sci 99:4149–4161PubMedGoogle Scholar
  55. 55.
    Hutchings RL, Singh SM, Cabello-Villegas J, Mallela KMG (2013) Effect of antimicrobial preservatives on partial protein unfolding and aggregation. J Pharm Sci 102:365–376PubMedGoogle Scholar
  56. 56.
    Bis RL, Singh SM, Cabello-villegas J, Mallela KMG (2014) Role of benzyl alcohol in the unfolding and aggregation of interferon alpha-2a. J Pharm Sci 26:1–9Google Scholar
  57. 57.
    Heljo P, Ross A, Zarraga IE, Pappenberger A, Mahler H-C (2015) Interactions between peptide and preservatives: effects on peptide self-interactions and antimicrobial efficiency in aqueous multi-dose formulations. Pharm Res 32:3201–3212PubMedGoogle Scholar
  58. 58.
    Jezek J et al (2013) Biopharmaceutical formulations for pre-filled delivery devices. Expert Opin Drug Deliv 10:811–828PubMedGoogle Scholar
  59. 59.
    Kocha T, Yamaguchi M, Ohtaki H, Fukuda T, Aoyagi T (1996) Hydrogen peroxide-mediated degradation of protein: different oxidation modes of copper- and iron-dependent hydroxyl radicals on the degradation of albumin. Biochim Biophys Acta 1337:319–326Google Scholar
  60. 60.
    Morefield GL et al (2005) Role of aluminum-containing adjuvants in antigen internalization by dendritic cells in vitro. Vaccine 23:1588–1595PubMedGoogle Scholar
  61. 61.
    Mbow ML, De Gregorio E, Valiante NM, Rappuoli R (2010) New adjuvants for human vaccines. Curr Opin Immunol 22:411–416PubMedGoogle Scholar
  62. 62.
    Guy B (2007) The perfect mix: recent progress in adjuvant research. Nat Rev Microbiol 5:505–517PubMedGoogle Scholar
  63. 63.
    Buonsanti C, Oro UD (2017) Discovery of immune potentiators as vaccine adjuvants. In: Immunopotentiators in modern vaccines. Elsevier, Amsterdam, pp 85–104Google Scholar
  64. 64.
    EMEA/CPMP (2004) Guideline on adjuvants in vaccines. 1–18Google Scholar
  65. 65.
    Jones LS et al (2005) Effects of adsorption to aluminum salt adjuvants on the structure and stability of model protein antigens. J Biol Chem 280:13406–13414PubMedGoogle Scholar
  66. 66.
    Fox CB, Kramer RM, Lucien Barnes V, Dowling QM, Vedvick TS (2013) Working together: interactions between vaccine antigens and adjuvants. Ther Adv Vaccines Rev 1:7–20Google Scholar
  67. 67.
    Kaurav M et al (2018) Combined adjuvant-delivery system for new generation vaccine antigens: alliance has its own advantage. Artif Cells Nanomed Biotechnol 46:S818–S831PubMedGoogle Scholar
  68. 68.
    Yanan C, Ping C, Binlong C, Suxin L, Hua G (2017) Monoclonal antibodies: formulations of marketed products and recent advances in novel delivery system. Drug Dev Ind Pharm 43:519–530Google Scholar
  69. 69.
    EPAR – Product Information. European Medicines Agency. Accessed 13 Oct 2019
  70. 70.
    Walsh G (2018) Biopharmaceutical benchmarks 2018. Nat Biotechnol 36:1136–1145PubMedGoogle Scholar
  71. 71.
    EMEA/CHMP (2015) Guideline on similar biological medicinal products. pp 1–7Google Scholar
  72. 72.
    Kirchhoff CF et al (2017) Biosimilars: key regulatory considerations and similarity assessment tools. Biotechnol Bioeng 114:2696–2705PubMedPubMedCentralGoogle Scholar
  73. 73.
    FDA (2015) Scientific considerations in demonstrating biosimilarity to a reference product. pp 1–24Google Scholar
  74. 74.
    EMEA/CHMP (2014) Guideline on similar biological medicinal products containing biotechnology-derived proteins as active substance: non-clinical and clinical issues. pp 1–13Google Scholar
  75. 75.
    Kesik-brodacka M (2018) Progress in biopharmaceutical development. Int Union Biochem Mol Biol 65:306–322Google Scholar
  76. 76.
    Anour R (2014) Biosimilars versus ‘biobetters’ – a regulator’s perspective. Generics Ans Biosimilars Initiat J 3:166–167Google Scholar
  77. 77.
    Ismael G et al (2012) Subcutaneous versus intravenous administration of (neo) adjuvant trastuzumab in patients with HER2-positive, clinical stage I–III breast cancer (HannaH study): a phase 3, open-label, multicentre, randomised trial. Lancet Oncol 13:869–878PubMedGoogle Scholar
  78. 78.
    Rathore N, Rajan RS (2008) Current perspectives on stability of protein drug products during formulation, fill and finish operations. Biotechnol Prog 24:504–514PubMedGoogle Scholar
  79. 79.
    Shnek DR, Hostettler DL, Bell MA, Olinger JM, Frank BH (1998) Physical stress testing of insulin suspensions and solutions. J Pharm Sci 87:1459–1465PubMedGoogle Scholar
  80. 80.
    Franks F (1998) Freeze-drying of bioproducts: putting principles into practice. Eur J Pharm Biopharm 45:221–229PubMedGoogle Scholar
  81. 81.
    Patel SM et al (2017) Lyophilized drug product cake appearance: what is acceptable? J Pharm Sci 106:1706–1721PubMedGoogle Scholar
  82. 82.
    Mensink MA, Frijlink HW, van der Voort K, Hinrichs WLJ (2017) How sugars protect proteins in the solid state and during drying (review): mechanisms of stabilization in relation to stress conditions. Eur J Pharm Biopharm 114:288–295PubMedGoogle Scholar
  83. 83.
    Cao W et al (2013) Rational design of lyophilized high concentration protein formulations-mitigating the challenge of slow reconstitution with multidisciplinary strategies. Eur J Pharm Biopharm 85:287–293PubMedGoogle Scholar
  84. 84.
    FDA (1999) Container closure systems for packaging human drugs and biologics. pp 1–41Google Scholar
  85. 85.
    Wang M et al (2018) Interactions between biological products and product packaging and potential approaches to overcome them. AAPS PharmSciTech 19:3681–3686PubMedGoogle Scholar
  86. 86.
    Raghani A, Li K, Bussiere JL, Bercu JP, Qiu J (2018) Process-related impurities in biopharmaceuticals. In: ICH quality guidelines: an implementation guide. Wiley, Hoboken, pp 487–507Google Scholar
  87. 87.
    Dipaolo B, Pennetti A, Nugent L, Venkat K, Venkat K (1999) Monitoring impurities in biopharmaceuticals produced by recombinant technology. Pharm Sci Technol Today 2:70–82PubMedGoogle Scholar
  88. 88.
    Florence AT, Attwood D (2016) Adverse events: the role of formulations and delivery systems. In: Physicochemical principles of pharmacy. Macmillan, Basingstoke, pp 481–511Google Scholar
  89. 89.
    Elder DP, Kuentz M, Holm R (2015) Pharmaceutical excipients – quality, regulatory and biopharmaceutical considerations. Eur J Pharm Sci 87:1–12Google Scholar
  90. 90.
    WHO (2013) Guidelines on the quality, safety, and efficacy of biotherapeutic protein products prepared by recombinant DNA technology. pp 1–91Google Scholar
  91. 91.
    Nally J (2007) Good manufacturing practices for pharmaceuticals. CRC Press, Boca RatonGoogle Scholar
  92. 92.
    EMEA/CHMP (2007) Guideline on excipients in the dossier for application for marketing authorization of a medical product. pp 1–12Google Scholar
  93. 93.
    Pichler WJ (2006) Adverse side-effects to biological agents. Allergy 61:912–920PubMedGoogle Scholar
  94. 94.
    Schellekens H (2002) Immunogenicity of therapeutic proteins: clinical implications and future prospects. Clin Ther 24:1720–1740PubMedGoogle Scholar
  95. 95.
    Tovey MG, Lallemand C (2011) Immunogenicity and other problems associated with the use of biopharmaceuticals. Ther Adv Drug Saf 2:113–128PubMedPubMedCentralGoogle Scholar
  96. 96.
    Schellekens H, Casadevall N (2004) Immunogenicity of recombinant human proteins: causes and consequences. J Neurol 251:4–9Google Scholar
  97. 97.
    Li J et al (2019) Thrombocytopenia caused by the development of antibodies to thrombopoietin. Am Soc Hematol 98:3241–3249Google Scholar
  98. 98.
    Schernthaner G (1993) Immunogenicity and allergenic potential of animal and human insulins. Diabetes Care 16:155–165PubMedGoogle Scholar
  99. 99.
    Dumont J, Euwart D, Mei B, Estes S, Kshirsagar R (2016) Human cell lines for biopharmaceutical manufacturing: history, status, and future perspectives. Crit Rev Biotechnol 36:1110–1122PubMedGoogle Scholar
  100. 100.
    Robinson AS (2012) Minimizing immunogenicity of biopharmaceuticals by controlling critical quality attributes of proteins. Biotechnol Bioeng 7:1473–1484Google Scholar
  101. 101.
    Hermeling S, Crommelin DJA, Schellekens H, Jiskoot W (2004) Structure-immunogenicity relationships of therapeutic proteins. Pharm Res 21:897–903Google Scholar
  102. 102.
    Kijanka G et al (2018) Submicron size particles of a murine monoclonal antibody are more immunogenic than soluble oligomers or micron size particles upon subcutaneous administration in mice. J Pharm Sci 107:2847–2859PubMedGoogle Scholar
  103. 103.
    Rosenberg AS (2006) Effects of protein aggregates: an immunologic perspective. AAPS J 8:501–507Google Scholar
  104. 104.
    The United States Pharmacopeial Convention (2012) <788> Particulate matter in injections. pp 1–3Google Scholar
  105. 105.
    European Pharmacopoeia (2007) 2.9.19. Particulate contamination: sub-visible particles. pp 300–302Google Scholar
  106. 106.
    European Pharmacopoeia (2007) 2.9.20. Particulate contamination: visible particles. p 302Google Scholar
  107. 107.
    Ito H, Nakashima T, So T, Hirata M, Inoue M (2003) Immunodominance of conformation-dependent B-cell epitopes of protein antigens. Biochem Biophys Res Commun 308:770–776PubMedGoogle Scholar
  108. 108.
    Casadevall N, Nataf J, Viron B, Kolta A, Patrick M (2002) Pure red cell aplasia and antierythropoietin antibodies in patients treated with recombinant erythropoietin. N Engl J Med 346:469–475PubMedGoogle Scholar
  109. 109.
    Shankar G, Pendley C, Stein KE (2007) A risk-based bioanalytical strategy for the assessment of antibody immune responses against biological drugs. Nat Biotechnol 25:555–561PubMedGoogle Scholar
  110. 110.
    Groot, A. S. De, Mcmurry, J. & Moise, L. Prediction of immunogenicity: in silico paradigms, ex vivo and in vivo correlates. Curr Opin Pharmacol 8, 1–7 (2008)Google Scholar
  111. 111.
    Shankar G et al (2014) Assessment and reporting of the clinical immunogenicity of therapeutic proteins and peptides – harmonized terminology and tactical recommendations. AAPS J 16:658–673PubMedPubMedCentralGoogle Scholar
  112. 112.
    EMEA/CHMP (2007) Guideline on immunogenicity assessment of biotechnology-derived therapeutic proteins. pp 1–18Google Scholar
  113. 113.
    EMEA/CHMP (2012) Guideline on immunogenicity assessment of monoclonal antibodies intended for in vivo clinical use. pp 1–10Google Scholar
  114. 114.
    FDA (2016) Assay development and validation for immunogenicity testing of therapeutic protein products. pp 1–31Google Scholar
  115. 115.
    Shah M (2018) New perspectives on protein aggregation during biopharmaceutical development. Int J Pharm 552:1–6PubMedGoogle Scholar
  116. 116.
    Bookbinder LH et al (2006) A recombinant human enzyme for enhanced interstitial transport of therapeutics. J Control Release 114:230–241PubMedGoogle Scholar
  117. 117.
    Schiefelbein L et al (2010) Synthesis, characterization and assessment of suitability of trehalose fatty acid esters as alternatives for polysorbates in protein formulation. Eur J Pharm Biopharm 76:342–350PubMedGoogle Scholar
  118. 118.
    Du W, Klibanov AM (2011) Hydrophobic salts markedly diminish viscosity of concentrated protein solutions. Biotechnol Bioeng 108:632–636PubMedGoogle Scholar
  119. 119.
    Serno T, Geidobler R, Winter G (2011) Protein stabilization by cyclodextrins in the liquid and dried state. Adv Drug Deliv Rev 63:1086–1106PubMedGoogle Scholar
  120. 120.
    Stella VJ, He Q (2008) Cyclodextrins. Toxicol Pathol 36:30–42PubMedGoogle Scholar
  121. 121.
    Kozarewicz P, Loftsson T (2018) Novel excipients – regulatory challenges and perspectives – the EU insight. Int J Pharm 546:176–179PubMedGoogle Scholar
  122. 122.
    FDA (2005) Nonclinical studies for the safety evaluation of pharmaceutical excipients. pp 1–9Google Scholar
  123. 123.
    Zhu L, Xu H (2015) The optimal choice of medication administration route regarding intravenous, intramuscular, and subcutaneous injection. Dove Press J 9:923–942Google Scholar
  124. 124.
    Fayad F et al (2018) Patient preferences for rheumatoid arthritis treatments: results from the national cross-sectional LERACS study. Dove Press J 12:1619–1625Google Scholar
  125. 125.
    Mahato RI, Narang AS, Th L, Miller DD (2003) Emerging trends in oral delivery of peptide and protein drugs. Crit Rev Ther Drug Carrier Syst 20:153–214PubMedGoogle Scholar
  126. 126.
    Barnett AH, Bellary S (2007) Inhaled human insulin (Exubera®): clinical profile and patient considerations. Vasc Health Risk Manag 3:83–91Google Scholar
  127. 127.
    Hanson LR, Ii WHF (2008) Intranasal delivery bypasses the blood-brain barrier to target therapeutic agents to the central nervous system and treat neurodegenerative disease. BMC Neurosci 9:1–4Google Scholar
  128. 128.
    Chaulagain B, Jain A, Tiwari A, Verma A, Jain SK (2018) Passive delivery of protein drugs through transdermal route. Artif Cells Nanomed Biotechnol 46:S472–S487Google Scholar
  129. 129.
    Ye Y, Yu J, Wen D, Kahkoska AR, Gu Z (2018) Polymeric microneedles for transdermal protein delivery. Adv Drug Deliv Rev 127:106–118PubMedPubMedCentralGoogle Scholar
  130. 130.
    Kayser O, Warzecha H (2012) Pharmaceutical biotechnology drug discovery and clinical applications. Wiley-Blackwell, HobokenGoogle Scholar
  131. 131.
    Philippart M et al (2016) Oral delivery of therapeutic proteins and peptides: an overview of current technologies and recommendations for bridging from approved intravenous or subcutaneous administration to novel oral regimens. Drug Res 66:113–120Google Scholar
  132. 132.
    Vaishya R, Khurana V, Patel S, Mitra AK (2015) Long-term delivery of protein therapeutics. Expert Opin Drug Deliv 12:415–440PubMedGoogle Scholar
  133. 133.
    Crommelin DJA, Hawe A, Jiskoot W (2019) Formulation of biologics including biopharmaceutical considerations. In: Pharmaceutical biotechnology. Springer, Berlin, pp 83–103Google Scholar
  134. 134.
    Ye C, Venkatraman S (2019) The long-term delivery of proteins and peptides using micro/nanoparticles: overview and perspectives. Ther Deliv 10:10–13Google Scholar
  135. 135.
    Vermonden T, Censi R, Hennink WE (2012) Hydrogels for protein delivery. Chem Rev 112:2853–2888PubMedGoogle Scholar
  136. 136.
    Capelle MAH, Arvinte T (2008) High-throughput formulation screening of therapeutic proteins. Drug Discov Today Technol 5:71–79Google Scholar
  137. 137.
    Senisterra GA, Finerty PJ (2009) High throughput methods of assessing protein stability and aggregation. Mol Biosyst 5:217–223PubMedGoogle Scholar
  138. 138.
    Carbonell F, Iturria-Medina Y, Evans AC (2018) Mathematical modeling of protein misfolding mechanisms in neurological diseases: a historical overview. Front Neurol 9:1–16Google Scholar
  139. 139.
    Andrews JM, Roberts CJ (2007) A Lumry-Eyring nucleated polymerization model of protein aggregation kinetics: 1.aggregation with pre-equilibrated unfolding. J Phys Chem 111:7897–7913Google Scholar
  140. 140.
    Mulder NJ, Kersey P, Pruess M, Apweiler R (2008) In silico characterization of proteins: UniProt, InterPro and Integr8. Mol Biotechnol 38:165–177PubMedGoogle Scholar
  141. 141.
    Kirchmair J et al (2008) The Protein Data Bank (PDB), its related services and software tools as key components for in silico guided drug discovery. J Med Chem 51:7021–7039PubMedGoogle Scholar
  142. 142.
    Tamizi E, Jouyban A (2016) Forced degradation studies of biopharmaceuticals: Selection of stress conditions. Eur J Pharm Biopharm 98:26–46PubMedGoogle Scholar
  143. 143.
    Nowak C et al (2017) Forced degradation of recombinant monoclonal antibodies: a practical guide. MAbs 9:1217–1230PubMedPubMedCentralGoogle Scholar
  144. 144.
    Hawe A et al (2012) Forced degradation of therapeutic proteins. J Pharm Sci 101:895–913PubMedGoogle Scholar
  145. 145.
    Capelle MAH, Gurny R, Arvinte T (2007) High throughput screening of protein formulation stability: practical considerations. Eur J Pharm Biopharm 65:131–148PubMedGoogle Scholar
  146. 146.
    Ducret A, Oostveen IVAN, Eng JK, In JRY, Aebersold R (1998) High throughput protein characterization by automated reverse-phase chromatography/electrospray tandem mass spectrometry. Protein Sci 7:706–719PubMedPubMedCentralGoogle Scholar
  147. 147.
    Zhao HUI et al (2010) Formulation development of antibodies using robotic system and high-throughput laboratory (HTL). J Pharm Sci 99:2279–2294PubMedGoogle Scholar
  148. 148.
    Wang W, Ohtake S (2019) Science and art of protein formulation development. Int J Pharm 568:118505PubMedGoogle Scholar
  149. 149.
    Perez-ramírez B, Guziewicz N, Simler R, Sreedhara A (2015) Approaches for early developability assessment of proteins to guide quality by design of liquid formulations. In: Quality by design for biopharmaceutical drug product development. Springer, Berlin, pp 87–114Google Scholar
  150. 150.
    Jarasch A et al (2015) Developability assessment during the selection of novel therapeutic antibodies. J Pharm Sci 104:1885–1898PubMedGoogle Scholar
  151. 151.
    Zurdo J (2013) Developability assessment as an early de-risking tool for biopharmaceutical development. Pharm Bioprocess 1:29–50Google Scholar
  152. 152.
    ICH (2009) ICH Q8(R2) – pharmaceutical development. pp 1–24Google Scholar
  153. 153.
    Grant Y, Matejtschuk P, Bird C, Wadhwa M, Dalby PA (2012) Freeze drying formulation using microscale and design of experiment approaches: a case study using granulocyte colony-stimulating factor. Biotechnol Lett 34:641–648PubMedGoogle Scholar
  154. 154.
    ICH (2005) ICH Q9 – quality risk management. pp 1–19Google Scholar
  155. 155.
    Moreton C (2009) Functionality and performance of excipients in a quality-by-design world. Am Pharm Rev:32–35Google Scholar

Copyright information

© Springer Nature Switzerland AG 2019

Authors and Affiliations

  • Rita Ribeiro
    • 1
    • 2
  • Teresa Raquel Abreu
    • 1
    • 2
  • Ana Catarina Silva
    • 3
    • 4
  • João Gonçalves
    • 5
  • João Nuno Moreira
    • 1
    • 2
    Email author
  1. 1.CNC – Center for Neuroscience and Cell Biology, Faculty of Medicine (Pólo I)University of CoimbraCoimbraPortugal
  2. 2.FFUC – Faculty of PharmacyUniversity of CoimbraCoimbraPortugal
  3. 3.UCIBIO, UCIBIO, REQUIMTE, MEDTECH, Laboratory of Pharmaceutical Technology, Department of Drug Sciences, Faculty of PharmacyUniversity of PortoPortoPortugal
  4. 4.FP-ENAS (UFP Energy, Environment and Health Research Unit), CEBIMED (Biomedical Research Centre), Faculty of Health SciencesUniversity Fernando PessoaPortoPortugal
  5. 5.iMed.ULisboa – Research Institute for Medicines, Faculty of PharmacyUniversity of LisbonLisbonPortugal

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