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Biomimetic Affinity Ligands for Immunoglobulins Based on the Multicomponent Ugi Reaction

  • Graziella El Khoury
  • Laura A. Rowe
  • Christopher R. LoweEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 800)

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.

Key words

Affinity Biomimetic ligands Combinatorial synthesis Protein purification Multi­component reaction Rational design Ugi reaction 

Notes

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).

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Graziella El Khoury
    • 1
  • Laura A. Rowe
    • 1
  • Christopher R. Lowe
    • 1
    Email author
  1. 1.Department of Chemical Engineering and BiotechnologyUniversity of CambridgeCambridgeUK

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