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Exploring variation in binding of Protein A and Protein G to immunoglobulin type G by isothermal titration calorimetry

Lund, Line Naomi; Christensen, Trine; Toone, Eric; Houen, Gunnar; Staby, Arne LU and St Hilaire, Phaedria Marie (2011) In Journal of Molecular Recognition 24(6). p.945-952
Abstract
Bacterial Protein A (PrtA) and Protein G (PrtG) are widely used for affinity purification of antibodies. An understanding of how PrtA and PrtG bind to different isotypes of immunoglobulin type G (IgG) and to their corresponding Fc fragments is essential for the development of PrtA and PrtG mimetic ligands and for the establishment of generic processes for the purification of various antibodies. In this paper, the interactions between the two IgG-binding proteins and IgG of two different subclasses, IgG1 and IgG4, as well as their analogous Fc fragments have been studied by isothermal titration calorimetry. The results indicate that both protein ligands bind IgG and Fc fragments strongly with Ka values in the range of 10(7)-10(8) M(-1) and... (More)
Bacterial Protein A (PrtA) and Protein G (PrtG) are widely used for affinity purification of antibodies. An understanding of how PrtA and PrtG bind to different isotypes of immunoglobulin type G (IgG) and to their corresponding Fc fragments is essential for the development of PrtA and PrtG mimetic ligands and for the establishment of generic processes for the purification of various antibodies. In this paper, the interactions between the two IgG-binding proteins and IgG of two different subclasses, IgG1 and IgG4, as well as their analogous Fc fragments have been studied by isothermal titration calorimetry. The results indicate that both protein ligands bind IgG and Fc fragments strongly with Ka values in the range of 10(7)-10(8) M(-1) and for both ligands, the interaction with both IgG isotypes is enthalpically driven though entropically unfavorable. Moreover, variation in the standard entropic and standard enthalpic contribution to binding between the two isotypes as well as between IgG and Fc fragment implies that the specific interaction with PrtA varies according to IgG isotype. In contrast to PrtA, PrtG bound to F(ab')(2) fragment with a Ka value of 5.1 x 10(5) M(-1); thus underscoring the usefulness of PrtA as a preferred ligand for generic antibody purification processes. Copyright (C) 2011 John Wiley & Sons, Ltd. (Less)
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organization
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Contribution to journal
publication status
published
subject
keywords
Protein A, Protein G, immunoglobulin G, Fc fragment, isothermal, titration calorimetry
in
Journal of Molecular Recognition
volume
24
issue
6
pages
945 - 952
publisher
John Wiley & Sons
external identifiers
  • wos:000298599500006
  • scopus:80155178893
ISSN
1099-1352
DOI
10.1002/jmr.1140
language
English
LU publication?
yes
id
5b72eebe-f070-4b06-b2de-fa0deed4c4c3 (old id 2313127)
date added to LUP
2012-02-07 09:56:11
date last changed
2017-08-27 03:17:17
@article{5b72eebe-f070-4b06-b2de-fa0deed4c4c3,
  abstract     = {Bacterial Protein A (PrtA) and Protein G (PrtG) are widely used for affinity purification of antibodies. An understanding of how PrtA and PrtG bind to different isotypes of immunoglobulin type G (IgG) and to their corresponding Fc fragments is essential for the development of PrtA and PrtG mimetic ligands and for the establishment of generic processes for the purification of various antibodies. In this paper, the interactions between the two IgG-binding proteins and IgG of two different subclasses, IgG1 and IgG4, as well as their analogous Fc fragments have been studied by isothermal titration calorimetry. The results indicate that both protein ligands bind IgG and Fc fragments strongly with Ka values in the range of 10(7)-10(8) M(-1) and for both ligands, the interaction with both IgG isotypes is enthalpically driven though entropically unfavorable. Moreover, variation in the standard entropic and standard enthalpic contribution to binding between the two isotypes as well as between IgG and Fc fragment implies that the specific interaction with PrtA varies according to IgG isotype. In contrast to PrtA, PrtG bound to F(ab')(2) fragment with a Ka value of 5.1 x 10(5) M(-1); thus underscoring the usefulness of PrtA as a preferred ligand for generic antibody purification processes. Copyright (C) 2011 John Wiley & Sons, Ltd.},
  author       = {Lund, Line Naomi and Christensen, Trine and Toone, Eric and Houen, Gunnar and Staby, Arne and St Hilaire, Phaedria Marie},
  issn         = {1099-1352},
  keyword      = {Protein A,Protein G,immunoglobulin G,Fc fragment,isothermal,titration calorimetry},
  language     = {eng},
  number       = {6},
  pages        = {945--952},
  publisher    = {John Wiley & Sons},
  series       = {Journal of Molecular Recognition},
  title        = {Exploring variation in binding of Protein A and Protein G to immunoglobulin type G by isothermal titration calorimetry},
  url          = {http://dx.doi.org/10.1002/jmr.1140},
  volume       = {24},
  year         = {2011},
}