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Fc domain mediated self-association of an IgG1 monoclonal antibody under a low ionic strength condition

Paper ID Volume ID Publish Year Pages File Format Full-Text
21073 43205 2011 7 PDF Available
Title
Fc domain mediated self-association of an IgG1 monoclonal antibody under a low ionic strength condition
Abstract

Recently, we reported that IgG1 monoclonal antibody A (MAb A) underwent liquid–liquid phase separation and separated into light and heavy phases under a low ionic strength condition. The liquid–liquid phase separation was induced due to self-association of MAb A in the heavy phase when the initial concentration of MAb A was between the two critical concentrations [Nishi et al., Pharm. Res., 27, 1348-1360 (2010)]. Here, we determined the interaction site of MAb A by using proteolytic Fab and Fc fragments of MAb A. The mean hydrodynamic diameter of the Fc fragment increased in a low ionic strength buffer, and furthermore the SPR measurement detected interactions of the Fc fragment with both whole MAb A and the Fc fragment, whereas the Fab fragment interacted with neither whole MAb A nor the Fc fragment. No binding was detected under an isotonic ionic strength condition. Zeta potential of MAb A was significant positive below pH 5.5 and negative above pH 6.5. Between pH 5.5 and 6.5 where the phase separation is significantly induced, MAb A had only a small positive or negative net charge. The isothermal titration calorimetry dilution method revealed that dissociation of MAb A accompanied endothermic heat changes, suggesting that intermolecular interactions among MAb A molecules were attributed to the enthalpically driven process. These results suggest that liquid–liquid phase separation of MAb A is mediated by a weak electrostatic intermolecular interaction among MAb A molecules mainly at Fc portions.

Keywords
Monoclonal antibody; Liquid–liquid phase separation; Protein self-association; Fc–Fc interaction; Low ionic strength condition; Dynamic light scattering (DLS); Surface plasmon resonance (SPR); Isothermal titration calorimetry (ITC)
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Fc domain mediated self-association of an IgG1 monoclonal antibody under a low ionic strength condition
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Bioscience and Bioengineering - Volume 112, Issue 4, October 2011, Pages 326–332
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us