Unbinding forces of single pertussis toxin–antibody complexes measured by atomic force spectroscopy correlate with their dissociation rates determined by surface plasmon resonance

Moreno, Nadège; Chevalier, Michel; Ronzon, Frédéric; Manin, Catherine; Dupuy, Monique; Krell, Tino; Rieu, Jean-Paul

doi:10.1002/jmr.1159

Cited by 11 publications

(11 citation statements)

References 48 publications

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“…S3 A, B). Previous AFM studies showed a satisfying correlation between the 2D off-rate extrapolated at zero force (k o ) and 3D off-rate as measured with SPR (34,35). However, our results show that Bell Force are strongly different: F k ∼100 pN for C21 corresponds to a potential width of 0.04 nm in the energy landscape, likely related to a stiff bond (36).…”

Section: Discussionsupporting

confidence: 60%

Nanobody-antigen catch-bond reveals NK cell mechanosensitivity

González

Chames

Kerfélec

et al. 2018

Preprint

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Antibodies are key tools in biomedical research and medicine. Their binding properties are classically measured in solution and characterized by an affinity. However, in physiological conditions, antibodies can bridge an immune effector cell and an antigen presenting cell, implying that mechanical forces apply to the bonds. For example, in antibody-dependent cell cytotoxicity, a major mode of action of therapeutic monoclonal antibodies, the Fab domains bind the antigens on the target cell, while the Fc domain binds to the activating receptor CD16 (also known as FcgRIII) of an immune effector cell, in a quasi bi-dimensional environment (2D). Therefore, there is a strong need to investigating antigen/antibody binding under force (2D), to better understand and predict antibody activity in vivo. We used two anti-CD16 nanobodies targeting two different epitopes and laminar flow chamber assay to measure the association and dissociation of single bonds formed between microsphere-bound CD16 antigens and surface-bound anti-CD16 nanobodies (or single domain antibodies), simulating 2D encounters. The two nanobodies exhibit similar 2D association kinetics, characterized by a strong dependence on the molecular encounter duration. However, their 2D dissociation kinetics strongly differ as a function of applied force: one exhibits a slip bond behaviour where off-rate increases with force; the other exhibits a catch bond behaviour with off-rate decreasing with force. This is the first time, to our knowledge, that catch bond behaviour was reported for antigen-antibody bond. We further exploit this property to show how Natural Killer cells spread differentially on surfaces coated with these molecules, revealing NK cells mechanosensitivity. Our results may also have strong implications for the design of efficient bispecific antibodies for therapeutic applications.

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Section: Discussionsupporting

confidence: 60%

Nanobody-antigen catch-bond reveals NK cell mechanosensitivity

González

Chames

Kerfélec

et al. 2018

Preprint

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“…found a good correlation between DFS unbinding forces and SPR dissociation rates. 60 However, for other pairs, the dissociation rates differ by a factor of 10, indicating different dissociation paths. Residence times may also contribute to the difference in dissociation rate since, in DFS experiments, the tip-bound ligand contacts the α IIb β 3 receptor for less than 30 s, which may be too short to complete the stabilization steps detected by SPR.…”

Section: Discussionmentioning

confidence: 99%

PROBING αIIbβ3: LIGAND INTERACTIONS BY DYNAMIC FORCE SPECTROSCOPY AND SURFACE PLASMON RESONANCE

et al. 2013

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The interaction between platelet integrin αIIbβ3 and fibrin(ogen) plays a key role in blood clot formation and stability. Integrin antagonists, a class of pharmaceuticals used to prevent and treat cardiovascular disease, are designed to competitively interfere with this process. However, the energetics of the integrin–drug binding are not fully understood, potentially hampering further development of this class of pharmaceuticals. We integrated dynamic force spectroscopy (DFS) and surface plasmon resonance (SPR) to probe the energetics of complex formation between αIIbβ3 and cHarGD, a cyclic peptide integrin antagonist. Analysis of αIIbβ3:cHarGD DFS rupture force data at pulling rates of 14 000 pN/s, 42 000 pN/s and 70 000 pN/s yielded koff = 0.02–0.09 s−1, a dissociation energy barrier ΔGd‡ = 22–29 kJ/mol, and a potential well width x−1 = 0.5–0.8 nm. SPR kinetic data yielded an association rate constant kon = 7 × 103 L/mol-s and a dissociation rate constant koff = 10−2 s−1, followed by a slower stabilization step (τ ~ 400 s). Both DFS and SPR detected minimal interactions between αIIbβ3 and cHarGA demonstrating a key role for electrostatic interactions between the ligand aspartate and the integrin metal ion-dependent adhesion site (MIDAS). Our work provides new insights into the energy landscape of αIIbβ3’s interactions with pharmacological and physiological ligands.

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“…Furthermore, both lectins showed a decrease in recognition with increased crosslinking, implying that the availability of galactose is reduced. Studies have shown that crosslinking alters the antibody to antigen interaction due to the obstruction of recognition sites [ 32 ]. Furthermore, K88 adhesins binding to MS-Lac treatments were comparable to plant lectins, where MS-Lac1 exhibited greater biorecognition.…”

Section: Discussionmentioning

confidence: 99%

K88 Fimbrial Adhesin Targeting of Microspheres Containing Gentamicin Made with Albumin Glycated with Lactose

Sarabia-Sainz

Montfort

et al. 2015

IJMS

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The formulation and characterization of gentamicin-loaded microspheres as a delivery system targeting enterotoxigenic Escherichia coli K88 (E. coli K88) was investigated. Glycated albumin with lactose (BSA-glucose-β (4-1) galactose) was used as the microsphere matrix (MS-Lac) and gentamicin included as the transported antibiotic. The proposed target strategy was that exposed galactoses of MS-Lac could be specifically recognized by E. coli K88 adhesins, and the delivery of gentamicin would inhibit bacterial growth. Lactosylated microspheres (MS-Lac1, MS-Lac2 and MS-Lac3) were obtained using a water-in-oil emulsion, containing gentamicin, followed by crosslinking with different concentrations of glutaraldehyde. Electron microscopy displayed spherical particles with a mean size of 10–17 µm. In vitro release of gentamicin from MS-Lac was best fitted to a first order model, and the antibacterial activity of encapsulated and free gentamicin was comparable. MS-Lac treatments were recognized by plant galactose-specific lectins from Ricinus communis and Sophora japonica and by E. coli K88 adhesins. Results indicate MS-Lac1, produced with 4.2 mg/mL of crosslinker, as the best treatment and that lactosylated microsphere are promising platforms to obtain an active, targeted system against E. coli K88 infections.

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Unbinding forces of single pertussis toxin–antibody complexes measured by atomic force spectroscopy correlate with their dissociation rates determined by surface plasmon resonance

Cited by 11 publications

References 48 publications

Nanobody-antigen catch-bond reveals NK cell mechanosensitivity

Nanobody-antigen catch-bond reveals NK cell mechanosensitivity

PROBING αIIbβ3: LIGAND INTERACTIONS BY DYNAMIC FORCE SPECTROSCOPY AND SURFACE PLASMON RESONANCE

K88 Fimbrial Adhesin Targeting of Microspheres Containing Gentamicin Made with Albumin Glycated with Lactose

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