Inhibition of weak-affinity epitope-IgE interactions prevents mast cell degranulation

Handlogten, Michael W.; Kiziltepe, Tanyel; Serezani, Ana Paula Moreira; Kaplan, Mark H.; Bilgiçer, Başar

doi:10.1038/nchembio.1358

Cited by 35 publications

(52 citation statements)

References 39 publications

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“…33–38 In multiple publications, we established that the UV-NBS method can be used to functionalize antibodies without unfavorably impacting their antigen binding activity. Furthermore, we also demonstrated many utilities that the NBS can provide in a variety of applications beyond covalent linking such as oriented immobilization, 35,36 enhanced antigen detection, 35,37,38 as well as inhibition of allergic reactions 39–41 .…”

Section: Introductionmentioning

confidence: 91%

Site-specific conjugation of an antibody on a gold nanoparticle surface for one-step diagnosis of prostate specific antigen with dynamic light scattering

2017

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Small dimensions of gold nanoparticles (AuNPs) necessitate antibodies to be immobilized in an oriented fashion in order to conserve their antigen binding activity for proper function. In this study, we used the previously described UV-NBS method to site-specifically incorporate thioctic acid (TA) functionality into antibodies at the conserved nucleotide-binding site (NBS). Modified antibodies were immobilized on AuNP surface in an oriented manner utilizing the newly incorporated TA functionality while maintaining antibody structure and activity. The resulting antibody functionalized AuNPs via the UV-NBS method demonstrated significantly enhanced antigen detection capabilities and improved antigen detection sensitivity with high level of selectivity when compared to other commonly used AuNP functionalization methods. Our results demonstrate that the limit of detection (LOD) for AuNPs functionalized via the UV-NBS method was 55 pM PSA, which is 40, 851, and 5873-fold improved over the other immobilization methods: EDC-NHS, thiol reduction, and ionic interaction, respectively. Consequently, the UV-NBS method provides a universal, site-specific functionalization method that generates highly sensitive and more stable antibody functionalized AuNPs that is amenable to any available detection and treatment assay with potential significant implications.

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Section: Introductionmentioning

confidence: 91%

Site-specific conjugation of an antibody on a gold nanoparticle surface for one-step diagnosis of prostate specific antigen with dynamic light scattering

2017

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“…In our previous work, we describe the design, synthesis, and characterization of synthetic tetravalent allergens (Handlogten et al, 2013; Handlogten et al, 2012; Handlogten et al, 2013). These well-defined tetravalent allergens have several advantages compared to the widely used haptenated proteins, such as DNP conjugated to BSA (DNP-BSA), as model allergens.…”

Section: Resultsmentioning

confidence: 99%

Two-Allergen Model Reveals Complex Relationship between IgE Crosslinking and Degranulation

Handlogten

Deak

Bilgiçer

2014

Chemistry & Biology

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Summary Allergy is an immune response to complex mixtures of multiple allergens yet current models use a single synthetic allergen. Multiple allergens were modeled using two well-defined tetravalent allergens each specific for a distinct IgE thus enabling a systematic approach to evaluate the effect of each allergen and percent of allergen specific IgE on mast cell degranulation. We found the overall degranulation response caused by two allergens is additive for low allergen concentrations or low percent specific IgE, does not change for moderate allergen concentrations with moderate to high percent specific IgE, and is reduced for high allergen concentrations with moderate to high percent specific IgE. These results provide further evidence that supra-optimal IgE cross-linking decreases the degranulation response and establishes the two allergen model as a relevant experimental system to elucidate mast cell degranulation mechanisms.

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“…23 These assays were run in triplicate and performed at least twice as independent experiments. Briefly, cells at 150,000 cells per mL per added to a cell culture 96 well plate (0.1 mL per well) and allowed to attach to the plate for 6 hours until plate was confluent.…”

Section: Methodsmentioning

confidence: 99%

“…17,21,25–27 This design allowed control over the avidity between the allergen molecule to receptor bound IgE’s. This system has been exceptionally valuable in studies of IgE-FcεRI clustering and enabled us to demonstrate the significance of weak affinity epitopes in triggering cellular degranulation.…”

Section: Introductionmentioning

confidence: 99%

Nanoallergens: A multivalent platform for studying and evaluating potency of allergen epitopes in cellular degranulation

Deak

Vrabel

Pizzuti

et al. 2016

Exp Biol Med (Maywood)

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Degranulation caused by type I hypersensitivity (allergies) is a complex biophysical process, and available experimental models for studying relevant immunoglobulin E (IgE) binding epitopes on allergen proteins lack the ability to adequately evaluate, rank and associate these epitopes individually and with each other. In this study, we propose a new allergy model system for studying potential allergen epitopes using nanoallergens, liposomes modified to effectively display IgE binding epitopes/haptens. By utilizing the covalently conjugated lipid tails on two hapten molecules (dinitrophenol and dansyl), hapten molecules were successfully incorporated into liposomes with high precision to form nanoallergens. Nanoallergens, with precisely controlled high particle valency, can trigger degranulation with much greater sensitivity than commonly used bovine serum albumin (BSA) conjugates. In Rat Basophil Leukemia (RBL) cell experiments, nanoallergens with only 2% hapten loading were able to trigger degranulation in vitro at concentrations as low as 10 pM. Additionally, unlike BSA-hapten conjugates, nanoallergens allow exact control over particle size and valency. By varying the nanoallergen parameters such as size, valency, monovalent affinity of hapten, and specific IgE ratios, we exposed the importance of these variables on degranulation intensity while demonstrating nanoallergens’ potential for evaluating both high and low affinity epitopes. The data presented in this article establish nanoallergen platform as a reliable and versatile allergy model to study and evaluate allergen epitopes in mast cell degranulation.

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Inhibition of weak-affinity epitope-IgE interactions prevents mast cell degranulation

Cited by 35 publications

References 39 publications

Site-specific conjugation of an antibody on a gold nanoparticle surface for one-step diagnosis of prostate specific antigen with dynamic light scattering

Site-specific conjugation of an antibody on a gold nanoparticle surface for one-step diagnosis of prostate specific antigen with dynamic light scattering

Two-Allergen Model Reveals Complex Relationship between IgE Crosslinking and Degranulation

Nanoallergens: A multivalent platform for studying and evaluating potency of allergen epitopes in cellular degranulation

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