Abstract
Affinity chromatography is the method of choice for biomolecule separation and isolation with highly specific target recognition; it is ideally suited to the purification of immunotherapeutic proteins (i.e., mAbs). Conventional affinity purification protocols are based on natural immunoglobulin (Ig)-binding proteins, which are expensive to produce, labile, unstable, and exhibit lot-to-lot variability. Biological ligands are now being replaced by cost-effective, synthetic ligands, derived from the concepts of rational design and combinatorial chemistry, aided by in silico approaches. In this chapter, we describe a new synthetic procedure for the development of affinity ligands for immunoglobulins based on the multicomponent Ugi reaction. The lead ligand developed herein is specific for the IgG-Fab fragment and mimics Protein L (PpL), an IgG-binding protein isolated from Peptostreptococcus magnus strains and usually used for the purification of antibodies and their fragments.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Nelson AL, Dhimolea E, Reichert JM (2010) Development trends for human monoclonal antibody therapeutics. Nat Rev Drug Discov 9: 767–774
Walsh G (2010) Biopharmaceutical benchmarks 2010. Nat Biotech 28: 917–926
Lowe CR, Dean PDG (1974) Affinity Chromatography. John Wiley and Sons, New York
Lowe CR, Lowe AR, Gupta G (2001) New developments in affinity chromatography with potential application in the production of biopharmaceuticals. J Biochem Biophys Methods 49: 561–574
Lowe CR, Burton SJ, Burton N et al (1990) New developments in affinity chromatography. J Mol Recognit 3: 117–122
Li RX, Dowd V, Stewart DJ et al (1998) Design, synthesis, and application of a Protein A mimetic. Nat Biotechnol 16: 190–195
Teng SF, Sproule K, Hussain A et al (1999) A strategy for the generation of biomimetic ligands for affinity chromatography. Combinatorial synthesis and biological evaluation of an IgG binding ligand. J Mol Recognit 12: 67–75
Teng SF, Sproule K, Hussain A et al (2000) Affinity chromatography on immobilized “biomimetic” ligands synthesis, immobilization and chromatographic assessment of an immunoglobulin G-binding ligand. J Chrom B 740: 1–15
Sproule K, Morrill P, Pearson JC et al (2000) New strategy for the design of ligands for the purification of pharmaceutical proteins by affinity chromatography. J Chrom B 740: 17–33
Filippusson H, Erlendsson LS, Lowe CR (2000) Design, synthesis and evaluation of biomimetic affinity ligands for elastases. J Mol Recognit 13: 370–81
Palanisamy UD, Winzor DJ, Lowe CR (2000) Synthesis and evaluation of affinity adsorbents for glycoproteins: an artificial lectin. J Chrom B 746: 265–281
Morrill PR, Gupta G, Sproule K et al (2002) Rational combinatorial chemistry-based selection, synthesis and evaluation of an affinity adsorbent for recombinant human clotting factor VII. J Chrom B 774: 1–15
Roque ACA, Lowe CR, Taipa AM (2005a) An artificial protein L for the purification of immunoglobulins and Fab fragments by affinity chromatography. J Chrom A 1064: 157–167
Roque ACA, Lowe CR, Taipa AM (2005b) Synthesis and screening of a rationally designed combinatorial library of affinity ligands mimicking protein L from Peptostreptococcus magnus. J Mol Recognit 18: 213–224
Lowe CR, Burton SJ, Pearson JC et al (1986) Design and application of bio-mimetic dyes in biotechnology. J Chrom 376: 121–130
Haigh JM (2008) Novel affinity ligands for immunoglobulins based on the multicomponent Ugi reaction. PhD thesis. University of Cambridge
Haigh JM, Hussain A, Mimmack MI et al (2009) Affinity ligands for immunoglobulins based on the multicomponent Ugi reaction. J Chrom B 877: 1440–1452
Veiderma M (2007) Chemistry of the isocyanides and their multicomponent reactions, including their libraries – the initatives of Ivar Ugi. Proc Estonian Acad Sci Chem 56: 98–102
Graille M, Stura EA, Housden NG et al (2001) Complex between Peptostreptococcus magnus protein L and a human antibody reveals structural convergence in the interaction modes of Fab binding proteins. Structure 9: 679–687
Graille M, Harrison S, Crump MP et al (2002) Evidence for plasticity and structural mimicry at the immunoglobulin light chain-protein L interface. J Biol Chem 277: 47500–47506
Porath I, Fornstedt N (1970) Group fractionation of plasma proteins on dipolar ion exchangers. J Chrom A 51: 479–489
Marcaccini S, Torroba T (2007) The use of the Ugi four-component condensation. Nat Protoc 2: 632–639
Sundberg L, Porath J (1974) Preparation of adsorbents for affinity chromatography: I. attachment of group-containing ligands to insoluble polymers by means of bifunctional oxiranes. J Chrom A 90: 87–98
Winzor D J (2004) Determination of binding constants by affinity chromatography. J Chrom A 1037: 351–367
Kasche V (2001) Physico-chemical boundaries in the continuous and one-step discontinuous affinity-chromatographic isolation of proteins J Biochem Biophys Methods 49: 49–62
Greene TW, Wuts PGM (1991) Protective groups in organic synthesis. John Wiley and Sons, New York
Acknowledgment
The authors would like to thank Dr. Jonathan M. Haigh for permission to reproduce figures from his PhD thesis at the University of Cambridge (2008).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
El Khoury, G., Rowe, L.A., Lowe, C.R. (2012). Biomimetic Affinity Ligands for Immunoglobulins Based on the Multicomponent Ugi Reaction. In: Zanders, E. (eds) Chemical Genomics and Proteomics. Methods in Molecular Biology, vol 800. Humana Press. https://doi.org/10.1007/978-1-61779-349-3_5
Download citation
DOI: https://doi.org/10.1007/978-1-61779-349-3_5
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-61779-348-6
Online ISBN: 978-1-61779-349-3
eBook Packages: Springer Protocols