Affinity matters for <scp>IgE</scp>‐blocking activity of allergen‐specific antibodies

Strobl, Maria; Demir, Hilal; Stadlmayr, Gerhard; Stracke, Florian; Hoelzl, R.; Bohle, Barbara; Wozniak‐Knopp, Gordana

doi:10.1111/all.15746

Cited by 8 publications

(6 citation statements)

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“…After confirming that the measurements of four mAbs with In view of the positive association of affinity and IgE-blocking activity 3 together with the requirement of several mAbs specific for different epitopes to successfully reduce allergic symptoms 7,22,23 we hypothesize that the repertoire of highly affine Mal d 1-specific antibodies induced by Bet v 1-SLIT might be more restricted than the repertoire induced by Mal d 1-SLIT. This may represent one reason for the limited curative success of AIT with birch pollen on the associated apple allergy in some individuals.…”

Section: Discussionsupporting

confidence: 61%

“…9 The Bet v 1-specific murine mAbs BIP1, BIP4 10 and mP16, 11 as well as the Mal d 1-specific human mAbs K1.1 and K1.1-1 were generated and characterized in an earlier publication. 3…”

Section: Recombinant Allergens and Allergen-specific Mabsmentioning

confidence: 99%

“…2 Using patient-derived human monoclonal antibodies (mAbs) we recently showed that IgE-blocking activity enhances with antibody affinity. 3 To better understand the mechanisms of successful AIT it would be interesting to assess the avidity, i.e. the net binding force of all specific antibodies in serum samples, in the course of therapy.…”

Section: Introductionmentioning

confidence: 99%

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Easy assessment of the avidity of polyclonal allergen‐specific serum antibodies

Strobl,

Demir,

Wozniak‐Knopp

et al. 2024

Clin Experimental Allergy

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IntroductionAllergen‐specific IgE‐blocking IgG antibodies contribute to successful allergen immunotherapy (AIT), however, not much is known about their affinity. Since affinity measurements of polyclonal antibodies in serum are technically challenging we evaluated the applicability of acidic disruption of antibody‐allergen complexes by a modified ELISA protocol with monoclonal antibodies (mAbs) specific for the relevant major allergens Betv1 and Mald1. Then, AIT‐induced blocking and non‐blocking Mald1‐specific antibodies in sera from individuals with or without reduced apple allergy were compared.MethodsAfter testing their pH stability coated recombinant allergens were incubated with (i) mAbs diluted in PBS or human serum and (ii) sera from individuals after sublingual administration of Mald1 or Betv1 for 16 weeks. Immune complexes were exposed to buffers in the pH range of 6.4–3.4 and residual antibodies were measured. Avidity indexes (AI), defined as the pH removing 50% of antibodies, were compared to the dissociation constants (KD) of mAbs determined by surface plasmon resonance.ResultsThe selected pH range was applicable to disrupt allergen complexes with highly affine mAbs without compromising allergen integrity. AIs of mAbs accorded with KD values and were unaffected by epitope specificity or the presence of serum proteins. The inter‐assay variability was <4% CV. Protective Mald1‐specific IgG antibodies from individuals with reduced apple allergy showed a higher collective binding strength than that of the non‐protective antibodies of individuals without reduced apple allergy.ConclusionAcidic disruption of allergen‐antibody complexes may be used to estimate the net‐binding force of polyclonal serum antibodies and eases the investigation of affinity‐related research questions in AIT.

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Section: Discussionsupporting

confidence: 61%

“…9 The Bet v 1-specific murine mAbs BIP1, BIP4 10 and mP16, 11 as well as the Mal d 1-specific human mAbs K1.1 and K1.1-1 were generated and characterized in an earlier publication. 3…”

Section: Recombinant Allergens and Allergen-specific Mabsmentioning

confidence: 99%

See 1 more Smart Citation

Easy assessment of the avidity of polyclonal allergen‐specific serum antibodies

Strobl,

Demir,

Wozniak‐Knopp

et al. 2024

Clin Experimental Allergy

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“…Apple extract was provided by Prof. Barbara Bohle and prepared as described ( 39 ). Monoclonal IgG antibodies, BIP 1 and BIP 3 were kindly provided by Prof. Barbara Bohle ( 40 ). Mouse anti-HA-tag antibody and donkey anti-rabbit antibody both labeled with HRP were obtained from Sigma-Aldrich (St. Louis, MO, USA).…”

Section: Methodsmentioning

confidence: 99%

“…Based on the results of the initial experiment, Bet v 1 (final concentrations: 50 ng/ml and 100 ng/ml; 5 µl) was pre-incubated for two hours at room temperature with either an excess (330 and 370 times, respectively; 10 µl) of Nb32 and Nb32ILZ, or with an excess (330 times; 10 µl) of BIP 1, a Bet v 1-specific mouse IgG antibody ( 40 ) or polyclonal Bet v 1-specific rabbit serum (diluted 1:1200 in RPMI 1640, 10µl) before incubation with patient´s serum (20 µl). For control purposes, Bet v 1 was pre-incubated with an unrelated nanobody (excess: 370 times, 10 µl), BIP 3, an unrelated mouse monoclonal IgG antibody (excess: 330 times, 10 µl) ( 40 ) or unrelated rabbit serum (diluted 1:1200 in RPMI 1640, 10 µl). Bet v 1-IgE complexes were added to EBV-transformed B cells (1 x 10 5 /vial, 15 µl) and processed as described above.…”

Section: Methodsmentioning

confidence: 99%

Trimeric Bet v 1-specific nanobodies cause strong suppression of IgE binding

Bauernfeind,

Zettl,

Ivanova

et al. 2024

Front. Immunol.

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BackgroundAround 20% of the population in Northern and Central Europe is affected by birch pollen allergy, with the major birch pollen allergen Bet v 1 as the main elicitor of allergic reactions. Together with its cross-reactive allergens from related trees and foods, Bet v 1 causes an impaired quality of life. Hence, new treatment strategies were elaborated, demonstrating the effectiveness of blocking IgG antibodies on Bet v 1-induced IgE-mediated reactions. A recent study provided evidence for the first time that Bet v 1-specific nanobodies reduce patients´ IgE binding to Bet v 1. In order to increase the potential to outcompete IgE recognition of Bet v 1 and to foster cross-reactivity and cross-protection, we developed Bet v 1-specific nanobody trimers and evaluated their capacity to suppress polyclonal IgE binding to corresponding allergens and allergen-induced basophil degranulation.MethodsNanobody trimers were engineered by adding isoleucine zippers, thus enabling trimeric formation. Trimers were analyzed for their cross-reactivity, binding kinetics to Bet v 1, and related allergens, and patients’ IgE inhibition potential. Finally, their efficacy to prevent basophil degranulation was investigated.ResultsTrimers showed enhanced recognition of cross-reactive allergens and increased efficiency to reduce IgE-allergen binding compared to nanobody monomers. Furthermore, trimers displayed slow dissociation rates from allergens and suppressed allergen-induced mediator release.ConclusionWe generated high-affine nanobody trimers that target Bet v 1 and related allergens. Trimers blocked IgE-allergen interaction by competing with IgE for allergen binding. They inhibited IgE-mediated release of biological mediators, demonstrating a promising potential to prevent allergic reactions caused by Bet v 1 and relatives.

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The role of IgG1 and IgG4 as dominant IgE-blocking antibodies during allergen immunotherapy

Bohle

2024

Allergo J

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Affinity matters for IgE‐blocking activity of allergen‐specific antibodies

Abstract: 7. Ji C, Huang Y, Yeung LH, et al. Ara h 2-specific IgE presence rather than its function is the best predictor of mast cell activation in children.

Cited by 8 publications

References 7 publications

Easy assessment of the avidity of polyclonal allergen‐specific serum antibodies

Easy assessment of the avidity of polyclonal allergen‐specific serum antibodies

Trimeric Bet v 1-specific nanobodies cause strong suppression of IgE binding

The role of IgG1 and IgG4 as dominant IgE-blocking antibodies during allergen immunotherapy

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