Monoclonal Antibodies against Birch Pollen Allergens: Characterization by Immunoblotting and Use for Single-Step Affinity Purification of the Major Allergen &lt;i&gt;Bet &lt;/i&gt;&lt;i&gt;v&lt;/i&gt; I

Jarolim, E.; Tejkl, Margarethe; Rohac, Madeleine; Schlerka, Gabriele; Scheiner, Otto; Kraft, Dietrich; Breitenbach, Michael; Rumpold, Helmut

doi:10.1159/000235000

Cited by 95 publications

(72 citation statements)

References 12 publications

(16 reference statements)

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“…2c). No cross reactivity of the rat anti-mouse IgE antibody with mouse IgG1 was detected using a monoclonal mouse IgG1 (BIP-1) [20] (data not shown).…”

Section: Allergen-specific Serum Antibody Levelsmentioning

confidence: 94%

Influence of the route of sensitization on local and systemic immune responses in a murine model of type I allergy

Repa

Wild²,

Hufnagl³

et al. 2004

Clinical and Experimental Immunology

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SUMMARYThe pathophysiological and immunological characteristics of allergic immune responses are controlled by a variety of factors. We have studied the extent to which the route of sensitization influences allergenspecific IgE synthesis and local airway inflammation using a mouse model of allergic sensitization to the major birch pollen allergen Bet v 1. Sensitization of BALB/c mice with recombinant (r)Bet v 1 was performed using intraperitoneal (IP), subcutaneous (SC) or aerosol (AS) sensitization protocols. Mice were analysed for allergen-specific serum antibodies by ELISA and IgE-dependent basophil degranulation. Proliferative responses and cytokine production of splenocytes were measured upon Bet v 1 stimulation in vitro . Bronchoalveolar lavages were performed after airway challenge with aerosolized birch pollen extract for assessment of eosinophilic airway inflammation and local cytokine production in vivo. Highest allergen specific IgE levels and IgE-dependent basophil degranulation were achieved using the SC route. High IL-5 production by spleen and lung cells was associated with pronounced eosinophilia in bronchoalveolar lavages. After IP sensitization, despite giving the highest IgG levels, only low IgE levels, basophil degranulation and IL-5 production were seen. On the other hand, AS sensitization, resulting in the lowest systemic IgE and IL-5 levels, led to a comparably strong airway inflammation as the SC route. Our finding that the route of sensitization can result in a dissociation of local and systemic immune responses may contribute to a better understanding of the pathogenesis of allergic diseases and help to develop new treatment strategies.Keywords type I allergy murine model sensitization route local immune response systemic immune response

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“…2c). No cross reactivity of the rat anti-mouse IgE antibody with mouse IgG1 was detected using a monoclonal mouse IgG1 (BIP-1) [20] (data not shown).…”

Section: Allergen-specific Serum Antibody Levelsmentioning

confidence: 94%

Influence of the route of sensitization on local and systemic immune responses in a murine model of type I allergy

Repa

Wild²,

Hufnagl³

et al. 2004

Clinical and Experimental Immunology

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“…Immunoblotting with a polyclonal rabbit antiserum raised against the S-layer protein of B. sphaericus CCM 2177 was performed as described previously (8). The presence of Bet v1 epitopes was checked by immunoreactivity with BIP1, a monoclonal mouse anti-Bet v1 antibody (15). The presence of Strep-tag I was checked by dot blot assays.…”

Section: Methodsmentioning

confidence: 99%

Molecular Characterization of the S-Layer Gene,sbpA, ofBacillus sphaericusCCM 2177 and Production of a Functional S-Layer Fusion Protein with the Ability To Recrystallize in a Defined Orientation while Presenting the Fused Allergen

Ilk

Völlenkle²,

Egelseer³

et al. 2002

Appl Environ Microbiol

115

185

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Crystalline bacterial cell surface layers (S-layers) represent the outermost cell envelope component of many bacteria and archaea (35,37,38). S-layers are composed of identical protein or glycoprotein subunits, and they completely cover the cell surface during all stages of bacterial growth and division. The S-layer subunits assemble into either oblique, square, or hexagonal lattices. In the case of Bacillaceae, the N-terminal part is involved in anchoring the S-layer subunits via a distinct type of secondary cell wall polymer (SCWP) to the rigid cell wall layer (2,5,21,25,34,35).S-layers are unique biomaterials with properties most relevant for applications in molecular nanotechnology, nanobiotechnology, and biomimetics (38). Many S-layer proteins recrystallize into regularly structured monolayers on solid supports, such as silicon wafers, gold chips, and silanized glass or plastic materials, as well as on Langmuir lipid films, on liposomes (20,23), and at the air-water interface. Pores passing through S-layer lattices are of identical size and morphology, and functional groups show a regular distribution and high density. For production of S-layer-based biosensors (38), affinity microparticles (45), and solid-phase immunoassays (3,4,39), functional groups in the S-layer lattice were exploited as covalent binding sites for biologically active macromolecules, such as enzymes, antibodies, or ligands. As alternatives to the existing technology, namely, immobilization by chemical methods, genetic approaches are particularly attractive for incorporation of functional peptide sequences into S-layer proteins, which must be done at positions that do not interfere with their self-assembly properties and their interaction with SCWP.

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“…SUMs with immobilized r Bet v 1a were incubated with solutions containing BIP 1 (8) in different concentrations in blocking buffer. After incubation for 60 min at 20°C and at least three washing steps, anti-mouse alkaline phosphatase conjugate (whole molecule, goat; Sigma-Aldrich; 1:1000 in blocking buffer) was transferred onto the SUM surface and incubated for 60 min.…”

Section: Studies On the Applicability Of Sums As Matrices For Immunoamentioning

confidence: 99%

2-D Protein Crystals as an Immobilization Matrix for Producing Reaction Zones in Dipstick-Style Immunoassays

Breitwieser¹,

Küpcü²,

Howorka³

et al. 1996

BioTechniques

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Monoclonal Antibodies against Birch Pollen Allergens: Characterization by Immunoblotting and Use for Single-Step Affinity Purification of the Major Allergen <i>Bet </i><i>v</i> I

Cited by 95 publications

References 12 publications

Influence of the route of sensitization on local and systemic immune responses in a murine model of type I allergy

Influence of the route of sensitization on local and systemic immune responses in a murine model of type I allergy

Molecular Characterization of the S-Layer Gene,sbpA, ofBacillus sphaericusCCM 2177 and Production of a Functional S-Layer Fusion Protein with the Ability To Recrystallize in a Defined Orientation while Presenting the Fused Allergen

2-D Protein Crystals as an Immobilization Matrix for Producing Reaction Zones in Dipstick-Style Immunoassays

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