Development and Characterization of Peptide Ligands of Immunoglobulin G Fc Region

Bioconjugate Chemistry
Nika KruljecTomaž Bratkovič

Abstract

Affinity chromatography based on bacterial immunoglobulin (Ig)-binding proteins represents the cornerstone of therapeutic antibody downstream processing. However, there is a pressing need for more robust affinity ligands that would withstand the harsh column sanitization conditions, while still displaying high selectivity for antibodies. Here, we report the development of linear peptide IgG ligands, identified from combinatorial phage-display library screens. The lead peptide was shown to compete with staphylococcal protein A for the IgG Fc region. Trimming analysis and alanine scanning revealed the minimal structural requirements of the peptide for Fc binding, and the minimized peptide GSYWYQVWF recognized all human IgG subtypes. Mutation of glutamine located at the nonessential position 6 to aspartate led to the optimized peptide GSYWYDVWF with 18-fold higher affinity ( KD app. 0.6 μM) compared to the parent peptide. When coupled to paramagnetic beads or a chromatographic matrix, the optimized ligand was shown to selectively enrich antibodies from complex protein mixtures.

References

Feb 26, 2000·Science·W L DeLanoJ A Wells
Apr 1, 1997·Chemical Reviews·George P. Smith, Valery A. Petrenko
May 26, 2005·Biochemical and Biophysical Research Communications·Tomaz BratkovicUros Urleb
Dec 6, 2005·Journal of Chromatography. a·Rainer HahnAlois Jungbauer
Dec 13, 2005·Analytical Biochemistry·Robert KarlssonDavid G Myszka
Feb 24, 2006·Journal of the American Chemical Society·Ricardo L A DiasJohn A Robinson
Oct 13, 2006·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Sophia HoberMartin Linhult
Oct 19, 2006·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Abhinav A ShuklaDuncan Low
Jan 2, 2009·Journal of Chromatography. a·Haiou YangRuben G Carbonell
Nov 21, 2009·The Journal of Physical Chemistry. B·D MoianiG Fassina
Jan 6, 2010·Journal of Molecular Recognition : JMR·Haiou YangRuben G Carbonell
Jan 15, 2011·Journal of Peptide Science : an Official Publication of the European Peptide Society·Helena Gradišar, Roman Jerala
Apr 26, 2011·New Biotechnology·John H Chon, Gregory Zarbis-Papastoitsis
Aug 5, 2011·Assay and Drug Development Technologies·Moran Jerabek-WillemsenStefan Duhr
Nov 1, 2011·Journal of Molecular Recognition : JMR·Francesca DinonGiorgio Fassina
Oct 31, 2012·Biotechnology and Bioengineering·Stefano MenegattiBalaji M Rao
Jan 27, 2015·Methods in Molecular Biology·Malgorzata E Kokoszka, Brian K Kay
Dec 6, 2016·Advanced Drug Delivery Reviews·Zehra ElgundiVeysel Kayser
Jul 12, 2017·Bioconjugate Chemistry·Nika Kruljec, Tomaž Bratkovič
Oct 1, 2014·Bioengineering·Petra GronemeyerJochen Strube
Jan 10, 2018·Bioengineering & Translational Medicine·Abhinav A ShuklaCarnley Norman

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Citations

Jan 2, 2020·International Journal of Molecular Sciences·Krištof Bozovičar, Tomaž Bratkovič
May 4, 2020·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Soledad L SaavedraMaría C Martínez Ceron
Jun 4, 2021·Scientific Reports·Krištof BozovičarTomaž Bratkovič
Aug 16, 2019·Life Sciences·Fatemeh Sadat Tabatabaei MirakabadJavad Ranjbari

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