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

Fei Shen, Orlando J. Rojas*, Jan Genzer, Patrick V. Gurgel, Ruben G. Carbonell

*Corresponding author for this work

    Research output: Contribution to journalArticleScientificpeer-review

    11 Citations (Scopus)

    Abstract

    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.

    Original languageEnglish
    Pages (from-to)1829-1841
    Number of pages13
    JournalANALYTICAL AND BIOANALYTICAL CHEMISTRY
    Volume408
    Issue number7
    DOIs
    Publication statusPublished - 2016
    MoE publication typeA1 Journal article-refereed

    Keywords

    • Peptide ligands
    • Immunoglobulin G
    • Peptide HWRGWV
    • Spacer arms
    • Quartz crystal microbalance
    • QUARTZ-CRYSTAL MICROBALANCE
    • SURFACE-PLASMON RESONANCE
    • STAPHYLOCOCCAL-ENTEROTOXIN-B
    • PROTEIN ADSORPTION
    • ANTIBODY IMMOBILIZATION
    • BIOMOLECULAR ADSORPTION
    • NANOFIBRILLAR CELLULOSE
    • VISCOELASTIC PROPERTIES
    • QUANTITATIVE-ANALYSIS
    • PORCINE PARVOVIRUS

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