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Titolo:
Engineering, purification and applications of His-tagged recombinant antibody fragments with specificity for the major birch pollen allergen, Bet v 1
Autore:
Flicker, S; Laffer, S; Steinberger, P; Alhani, B; Zhu, Y; Laukkanen, ML; Keinanen, K; Kraft, D; Valenta, R;
Indirizzi:
Univ Vienna, Inst Gen & Expt Pathol, AKH, A-1090 Vienna, Austria Univ Vienna Vienna Austria A-1090 pt Pathol, AKH, A-1090 Vienna, Austria VTT Biotechnol & Food Res, FIN-02044 Espoo, Finland VTT Biotechnol & Food Res Espoo Finland FIN-02044 N-02044 Espoo, Finland
Titolo Testata:
BIOLOGICAL CHEMISTRY
fascicolo: 1, volume: 381, anno: 2000,
pagine: 39 - 47
SICI:
1431-6730(200001)381:1<39:EPAAOH>2.0.ZU;2-H
Fonte:
ISI
Lingua:
ENG
Soggetto:
ESCHERICHIA-COLI; MONOCLONAL-ANTIBODY; BET-V-1; IGE; TREE; QUANTIFICATION; DIAGNOSIS; IDENTIFICATION; EXPRESSION; VERRUCOSA;
Keywords:
allergen; recombinant antibody fragment; standardization; type I allergy;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
45
Recensione:
Indirizzi per estratti:
Indirizzo: Valenta, R Univ Vienna, Inst Gen & Expt Pathol, AKH, Wahringer Gurtel 18-20, A-1090 Vienna, Austria Univ Vienna Wahringer Gurtel 18-20 Vienna Austria A-1090 stria
Citazione:
S. Flicker et al., "Engineering, purification and applications of His-tagged recombinant antibody fragments with specificity for the major birch pollen allergen, Bet v 1", BIOL CHEM, 381(1), 2000, pp. 39-47

Abstract

Type I allergy, an immunodisorder affecting almost 20% of the population worldwide, is based on the production of IgE antibodies against per se harmless allergens. We report the expression of hexahistidine-tagged antibody fragments (Fabs) with specificity for Bet v1, the major birch pollen allergen, in Escherichia con. The cDNA coding for the heavy chain fragment of a mouse monoclonal anti-Bet v1 antibody, Bip 1, was engineered by PCR to containa hexahistidine-encoding 3' end. The modified Bip1 heavy chain cDNA was co-expressed in E. coli XL-1 Blue with the Bip1 light chain cDNA using the combinatorial plasmid pComb3H. His-tagged recombinant (r) Bip1 Fabs were isolated by nickel affinity chromatography and rBip 1 Fabs without His-tag werepurified via affinity to rBet v1. rBip1 Fabs with and without His-tag bound specifically to rBet v 1 and, like Bet v1-specific human serum IgE and rabbit-anti rBet v1 antibodies, cross-reacted with Betv1-related allergens inother plant-species (alder, oak, hazelnut). We demonstrate the usefulness of His-tagged rBip 1 Fabs (1) for the identification of pollen samples containing Betv1 by particle blotting, (2) for the detection of Bet v1-specificIgE antibodies in human serum samples by sandwich ELISA and (3) for the quantification of Bet v1 in solution. Based on these examples we suggest to use rBip 1 Fabs for the detection of Bet v1 and Bet v1-related allergens in natural allergen sources for allergy prevention, as well as for the standardization of natural allergen extracts produced for diagnosis and immunotherapy of birch pollen allergy.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 05/12/20 alle ore 13:07:55