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Affinity interactions of human immunoglobulin G with short peptides: role of ligand spacer on binding, kinetics, and mass transfer


The interaction affinity between human IgG and a short peptide ligand (hexameric HWRGWV) was investigated by following the shifts in frequency and energy dissipation in a quartz crystal microbalance (QCM). HWRGWV was immobilized by means of poly(ethylene glycol) tethered on QCM sensors coated with silicon oxide, which enhanced the accessibility of the peptide to hIgG and also passivated the surface. Ellipsometry and ToF-SIMS were employed for surface characterization. The peptide ligand density was optimized to 0.88 chains nm−2, which enabled the interaction of each hIgG molecule with at least one ligand. The maximum binding capacity was found to be 4.6 mg m−2, corresponding to a monolayer of hIgG, similar to the values for chromatographic resins. Dissociation constants were lower than those obtained from resins, possibly due to overestimation of bound mass by QCM. Equilibrium thermodynamic and kinetic parameters were determined, shedding light on interfacial effects important for detection and bioseparation.

The interaction affinity between human IgG and a short peptide ligand was investigated by using quartz crystal microgravimetry, ellipsometry and ToF-SIMS. Equilibrium thermodynamic and kinetics parameters were determined, shedding light on interfacial effects important for detection and bioseparation.

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The authors acknowledge the financial support from Pathogen Removal and Diagnostic Technologies (PRDT), a subsidiary of ProMetic BioSciences Inc. O.J.R. is grateful for funding support by the Academy of Finland through its Center of Excellence Program (2014–2019) “Molecular Engineering of Biosynthetic Hybrid Materials Research” (HYBER).

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Correspondence to Orlando J. Rojas.

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Published in the topical collection Analytical Applications of Biomimetic Recognition Elements with guest editors Maria C. Moreno-Bondi and Elena Benito-Peña.

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Shen, F., Rojas, O.J., Genzer, J. et al. Affinity interactions of human immunoglobulin G with short peptides: role of ligand spacer on binding, kinetics, and mass transfer. Anal Bioanal Chem 408, 1829–1841 (2016).

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  • Peptide ligands
  • Immunoglobulin G
  • Peptide HWRGWV
  • Spacer arms
  • Quartz crystal microbalance