Induction of Polyclonal and Monoclonal Antibodies with a Single Allergen Separated from Birch Pollen Extract by SDS-PAGE and Nitrocellulose Blots

Wheeler, A. W.; Deards, M.J.; Fox, B.; Hickman, B.E.; Spackman, V.M.

doi:10.1159/000235057

Cited by 5 publications

(1 citation statement)

References 7 publications

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“…A number of mAb have been produced already against grass [12][13][14][15], ragweed [16], cat and dog [17] and mite [18] for this purpose. Recently, anti-Bet v I mAb have been also produced and used to purify the molecule [2,19]. In this work, several anti-Bet v I mAb were produced and characterized: they were used to define the epitopic structure of the molecule and identify the allergenic epitopes.…”

Section: Introductionmentioning

confidence: 99%

Mapping of Bet v I Epitopes by Using Murine Monoclonal Antibodies

Marc-Series¹,

Boutin²,

Vrancken³

et al. 1990

Int Arch Allergy Immunol

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The different determinants of birch pollen extracts, as shown by SDS-PAGE analysis, range from 10 to 94 kDa. These determinants were then electrotransferred on nitrocellulose strips and allowed to react with human IgE Ab from sensitive patients in order to identify the allergenic determinants. Several minor (43, 35, 28 and 21 kDa) and the major (17 kDa) allergenic determinants were identified. Murine monoclonal antibodies (mAb) were then produced against the major allergenic determinant (Bet v I) and their specificity confirmed by immunoblot. One of them, mAb 3F10, was used to affinity-purify the Bet v I. The purity of this material was confirmed by SDS-PAGE analysis and its reactivity on immunoblot against human IgE ensured its biological activity. These mAb were then gathered on four families based on their pattern of reactivity with Bet v I. Indeed four different epitopes on the molecule were identified. Binding inhibition studies using two of them (mAb 5F9 and 8F12) suggested that the epitopes of Bet v I recognized by these mAb are not overlapping. On another hand, the binding of 8H7 and 3F10 was partially inhibited by 5F9 and the binding of 3F10, by 8F12. These data suggest that those two latter epitopes are somewhat overlapping. Finally, the mAb 5F9 could inhibit the binding of human IgE on the affinity-purified Bet v I up to 40% and then shares a common idiotope with human specific IgE Ab of allergic patients.

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Section: Introductionmentioning

confidence: 99%

Mapping of Bet v I Epitopes by Using Murine Monoclonal Antibodies

Marc-Series¹,

Boutin²,

Vrancken³

et al. 1990

Int Arch Allergy Immunol

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Specificity mapping of patients IgE response towards the tree pollen major allergens Aln g I, Bet v I and Cor a I

Ipsen

Petersen

et al. 1992

Clin Experimental Allergy

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The specificity pattern of IgE from non-treated tree pollen allergic patients (n = 38) were evaluated by solid phase absorption of serum samples followed by CRIE on alder, birch and hazel CIE precipitation profiles. The majority of the serum samples seemed to contain IgE antibodies with the following characteristics: specific towards Bet v I alone and common between Aln g I, Bet v I and/or Cor a I, 'II'. The IgE specificity profiles observed for 95% of the sera tested are compatible with birch pollen allergens being the only sensitizing allergens, indicating that the patients react to allergens from other tree pollens of the Fagales order due to IgE cross-reaction with the major allergens of birch and alder and/or hazel pollens.

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Analytic aspects of the standardization of allergenic extracts

Ree

1997

Allergy

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Induction of Polyclonal and Monoclonal Antibodies with a Single Allergen Separated from Birch Pollen Extract by SDS-PAGE and Nitrocellulose Blots

Cited by 5 publications

References 7 publications

Mapping of Bet v I Epitopes by Using Murine Monoclonal Antibodies

Mapping of Bet v I Epitopes by Using Murine Monoclonal Antibodies

Specificity mapping of patients IgE response towards the tree pollen major allergens Aln g I, Bet v I and Cor a I

Analytic aspects of the standardization of allergenic extracts

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