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Synthetic Ligand Affinity Chromatography Purification of Human Serum Albumin and Related Fusion Proteins

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Part of the Methods in Molecular Biology book series (MIMB,volume 2178)

Abstract

Synthetic ligand affinity adsorbents offer an efficient means for purification of biopharmaceuticals. Single-isomer textile dye C.I. Reactive Blue and newer ligands developed by rational design and screening of chemical combinatorial libraries based on a triazine scaffold are routinely used for the capture and purification of these proteins from engineered recombinant expression systems. Here, we describe methods for the purification of recombinant human serum albumin and related fusion proteins using synthetic ligand affinity adsorbents.

Key words

  • Human serum albumin
  • Albumin fusion proteins
  • Affinity chromatography
  • Synthetic affinity adsorbents

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  • DOI: 10.1007/978-1-0716-0775-6_11
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References

  1. Cohn EJ, Strong LE, Hughes WL Jr et al (1946) Preparation and properties of serum and plasma proteins. IV. A system for the separation into fractions of the protein and lipoprotein components of biological tissues and fluids. J Am Chem Soc 68:459–475

    CrossRef  CAS  PubMed  Google Scholar 

  2. Burnouf T (2000) Plasma protein technologies; what next? Transfus Today 14:11–13

    Google Scholar 

  3. Bergloff JH, Eriksson S, Suomela H et al (1983) Albumin from human plasma, preparation and in vitro properties. In: Curling JM (ed) Separation of plasma proteins. Pharmacia, Uppsala, pp 51–58

    Google Scholar 

  4. Curling JM (1980) Albumin purification by ion-exchange chromatography. In: Curling JM (ed) Methods in plasma fractionation. Academic Press, London, pp 77–92

    Google Scholar 

  5. Stoltz JF, Rivat C, Gescheir C et al (1991) Chromatographic purification of human albumin for clinical use. Pharm Tech Int 3:60–65

    Google Scholar 

  6. Travis J, Bowen J, Tewkesbury D et al (1976) Isolation of albumin from human plasma and fractionation of albumin-depleted plasma. Biochem J 157:301–306

    CrossRef  CAS  PubMed  Google Scholar 

  7. Ghoggeri GM, Candidiano G, Delfino G et al (1985) Highly selective one-step chromatography of serum and urinary albumin on immobilized Cibacron Blue F3G-A: studies on normal and glycosylated albumin. Clin Chem Acta 145:205–211

    CrossRef  Google Scholar 

  8. Subramanian S (1984) Dye-ligand affinity chromatography; the interaction of Cibacron Blue F3GA with proteins and enzymes. CRC Crit Rev Biochem 16(2):169–205

    CrossRef  CAS  Google Scholar 

  9. Burton SJ (1992) Dye-ligand affinity chromatography. Methods Mol Biol 11:91–103

    CAS  PubMed  Google Scholar 

  10. Dean PD, Watson DH (1979) Protein purification using immobilised triazine dyes. J Chromatogr 165(3):301–319

    CrossRef  CAS  PubMed  Google Scholar 

  11. Leatherbarrow RJ, Dean PD (1980) Studies on the mechanism serum albumins binding to immobilized Cibacron Blue F3G A. Biochem J 189:27–34

    CrossRef  CAS  PubMed  Google Scholar 

  12. Metcalf EC, Crow B, Dean PD (1981) The effect of ligand pre-saturation on the interaction of serum albumins with immobilized Cibacron Blue 3G-A studied by affinity gel electrophoresis. Biochem J 199:465–472

    CrossRef  CAS  PubMed  Google Scholar 

  13. Hanggi D, Carr P (1985) Analytical evaluation of the purity of commercial preparations of Cibacron Blue F3GA and related dyes. Anal Biochem 149(1):91–104

    CrossRef  CAS  PubMed  Google Scholar 

  14. Burton SJ, McLoughlin SB, Stead CV et al (1988) Design and applications of biomimetic anthraquinone dyes. I. Synthesis and characterization of terminal ring isomers of C.I. Reactive Blue 2. J Chromatogr A 435:127–137

    CrossRef  CAS  Google Scholar 

  15. Scoble J, Scopes R (1996) Well Defined dye adsorbents for protein purification. J Mol Recognit 9:728–732

    CrossRef  CAS  PubMed  Google Scholar 

  16. Lowe CR (2001) Combinatorial approaches to affinity chromatography. Curr Opin Chem Biol 5(3):248–256

    CrossRef  CAS  PubMed  Google Scholar 

  17. Curling J (2004) Affinity chromatography – from textile dyes to synthetic ligands by design. BioPharm Int 17(7):34–42

    CAS  Google Scholar 

  18. Subramanian GM, Fiscella M, Lamouse-Smith A et al (2007) Albinterferon alpha-2b: a genetic fusion protein for the treatment of chronic hepatitis C. Nat Biotechnol 25:1411–1419

    CrossRef  CAS  PubMed  Google Scholar 

  19. Sheffield WP, Eltringham-Smith LJ (2011) Incorporation of albumin fusion proteins into fibrin clots in vitro and in vivo: comparison of difference fusion motifs recognized by factor XIIIa. BMC Biotechnol 11:127

    CrossRef  CAS  PubMed  Google Scholar 

  20. Melder RJ, Osborn BL, Riccobone T et al (2004) Pharmacokinetics and in vitro and in vivo anti-tumor response of an interleukin-2-human serum albumin fusion protein in mice. Cancer Immunol Immunother 54(6):535–547

    CrossRef  CAS  PubMed  Google Scholar 

  21. Osborn BL, Sekut L, Corcoran M et al (2002) Albutropin: a growth hormone-albumin fusion with improved pharmacokinetics and pharmacodynamics in rats and monkeys. Eur J Pharmacol 456:149–148

    CrossRef  CAS  PubMed  Google Scholar 

  22. Duttaroy A, Kanakaraj P, Osborn BL et al (2005) Development of a long-acting insulin analog using albumin fusion technology. Diabetes 54:251–258

    CrossRef  CAS  PubMed  Google Scholar 

  23. Gao Z, Bai G, Chen J et al (2009) Development, characterization, and evaluation of a fusion protein of a novel glucagon-like peptide-1 (GLP-1) analog and human serum albumin in Pichia pastoris. Biosci Biotechnol Biochem 73(3):688–694

    CrossRef  CAS  PubMed  Google Scholar 

  24. Parviainen MT, Harmoinen A, Jokela H (1985) Serum albumin assay with bromocresol purple dye. Scand J Clin Lab Invest 45:561–564

    CrossRef  CAS  PubMed  Google Scholar 

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Acknowledgments

Bastiaan Lobbezoo and Steve Burton for valuable discussions.

Prometic Bioseparations Ltd. is a trademark of Prometic Bioseparations Ltd. Mimetic Blue® is registered with the US Patent and Trademark Office. AlbuPure® and Albufuse® are registered trademarks of Albumedix Ltd. AlbuPure® is sold under license for the use in the purification of albumin fusion proteins with no warranty of merchantability or fitness for a particular purpose.

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Correspondence to Dev Baines .

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Williams, S., Morton, P., Baines, D. (2021). Synthetic Ligand Affinity Chromatography Purification of Human Serum Albumin and Related Fusion Proteins. In: Labrou, N.E. (eds) Protein Downstream Processing. Methods in Molecular Biology, vol 2178. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0775-6_11

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  • DOI: https://doi.org/10.1007/978-1-0716-0775-6_11

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0774-9

  • Online ISBN: 978-1-0716-0775-6

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