Abstract
Small recombinant antibody formats are rapidly cleared from circulation. The coupling of these molecules to human serum albumin (HSA) is an efficient strategy to prolong their in vivo half-life. Two methods are described in this chapter for the attachment of HSA to small antibody molecules: the direct fusion of HSA and the indirect coupling by the fusion of an albumin-binding domain to the antibody. Furthermore, an assay is described to verify the functionality of the albumin-binding domain, and a method is proposed to determine the antibodies serum half-life in mice.
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References
Beckman RA, Weiner LM, Davis HM (2007) Antibody constructs in cancer therapy: protein engineering strategies to improve exposure in solid tumors. Cancer 109(2):170–179
Chapman AP (2002) PEGylated antibodies and antibody fragments for improved therapy: a review. Adv Drug Deliv Rev 54(4):531–545
Dennis MS et al (2002) Albumin binding as a general strategy for improving the pharmacokinetics of proteins. J Biol Chem 277(38):35035–35043
Hamidi M, Azadi A, Rafiei P (2006) Pharmacokinetic consequences of pegylation. Drug Deliv 13(6):399–409
Holliger P, Hudson PJ (2005) Engineered antibody fragments and the rise of single domains. Nat Biotechnol 23(9):1126–1136
Holt LJ et al (2008) Anti-serum albumin domain antibodies for extending the half-lives of short lived drugs. Protein Eng Des Sel 21(5):283–288
Johansson MU et al (2002) Structure, specificity, and mode of interaction for bacterial albumin-binding modules. J Biol Chem 277(10):8114–8120
Jonsson A et al (2008) Engineering of a femtomolar affinity binding protein to human serum albumin. Protein Eng Des Sel 21(8):515–527
Kim SJ, Park Y, Hong HJ (2005) Antibody engineering for the development of therapeutic antibodies. Mol Cells 20(1):17–29
Kim J et al (2006) Albumin turnover: FcRn-mediated recycling saves as much albumin from degradation as the liver produces. Am J Physiol Gastrointest Liver Physiol 290(2):G352–G360
Kontermann RE (2005) Recombinant bispecific antibodies for cancer therapy. Acta Pharmacol Sin 26(1):1–9
Kontermann RE (2009) Strategies to extend plasma half-lives of recombinant antibodies. BioDrugs 23:93–109
Kraulis PJ et al (1996) The serum albumin-binding domain of streptococcal protein G is a three-helical bundle: a heteronuclear NMR study. FEBS Lett 378(2):190–194
Lencer WI, Blumberg RS (2005) A passionate kiss, then run: exocytosis and recycling of IgG by FcRn. Trends Cell Biol 15(1):5–9
Linhult M et al (2002) Mutational analysis of the interaction between albumin-binding domain from streptococcal protein G and human serum albumin. Protein Sci 11(2):206–213
Muller D et al (2007) Improved Pharmacokinetics of Recombinant Bispecific Antibody Molecules by Fusion to Human Serum Albumin. J Biol Chem 282(17):12650–12660
Nguyen A et al (2006) The pharmacokinetics of an albumin-binding Fab (AB.Fab) can be modulated as a function of affinity for albumin. Protein Eng Des Sel 19(7):291–297
Schlapschy M et al (2007) Fusion of a recombinant antibody fragment with a homo-amino-acid polymer: effects on biophysical properties and prolonged plasma half-life. Protein Eng Des Sel 20(6):273–284
Stork R, Muller D, Kontermann RE (2007) A novel tri-functional antibody fusion protein with improved pharmacokinetic properties generated by fusing a bispecific single-chain diabody with an albumin-binding domain from streptococcal protein G. Protein Eng Des Sel 20(11):569–576
Stork R et al (2008) N-glycosylation as novel strategy to improve pharmacokinetic properties of bispecific single-chain diabodies. J Biol Chem 283(12):7804–7812
Subramanian GM et al (2007) Albinterferon alpha-2b: a genetic fusion protein for the treatment of chronic hepatitis C. Nat Biotechnol 25(12):1411–1419
Weir AN et al (2002) Formatting antibody fragments to mediate specific therapeutic functions. Biochem Soc Trans 30(4):512–516
Yazaki PJ et al (2008) Biodistribution and tumor imaging of an anti-CEA single-chain antibody-albumin fusion protein. Nucl Med Biol 35(2):151–158
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Stork, R. (2010). Fusion Proteins with Improved PK. In: Kontermann, R., Dübel, S. (eds) Antibody Engineering. Springer Protocols Handbooks. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-01147-4_16
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DOI: https://doi.org/10.1007/978-3-642-01147-4_16
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