ISOLATION AND CHARACTERIZATION OF A LOW MOLECULAR WEIGHT ALLERGEN FRACTION OF Blomia tropicalis.

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Bibliographic Details
Title:	ISOLATION AND CHARACTERIZATION OF A LOW MOLECULAR WEIGHT ALLERGEN FRACTION OF Blomia tropicalis.
Authors:	Labrada, A.¹, Labrada, M.¹ labrada@biocen.colombus.cu, Uyema, K.¹, Navarro, B.¹, Febles, M.¹, Puerta, L.²
Source:	Revista VacciMonitor (Vacunología y Temas Afines). Oct2002, Vol. 11 Issue 4, p1. 1p.
Subjects:	ALLERGENS, MOLECULAR weights, MITES, ALLERGIES, RECOMBINANT DNA, NF-kappa B, AMINO acid sequence
Abstract:	The domestic mite Blomia tropicalis (Bt) is an important source of allergens causing allergic diseases in tropical countries. More than 20 allergens have been detected in the extract of Bt. By using the rDNA technology, 3 allergens with similar Molecular Weight around 14kD have been obtained and characterized; one of them (Blo-t-5) is thought to be a major allergen. Nevertheless, the native analogs of these recombinant proteins have not been isolated nor identified. The aim of the present work was to isolate the native low molecular allergens (14-17kD) of Bt and to characterize their IgE-binding activity. A partially purified preparation (LMWF) was obtained by precipitation of the freeze-dried allergen extract in 50% ammonium sulfate, and subsequent gelfiltration of the supernatant in Sephadex-G50. The analysis by SDS-PAGE in non-reducing conditions revealed a major 14kD band, which was resolved in reducing conditions, in two close bands at 16-17kD. This band accounted for 80% of the total protein content. The UV absorbance spectrum peaked at 214 and 256nm. The LMWF was able to inhibit up to 69% of total IgE-binding activity of the extract, as measured by IgE-inhibition ELISA, using a serum pool of allergic patients to Bt. Up to 60% IgE reactivity was found by ELISA, to tested sera from 42 allergic subjects. The Western Blotting analysis of selected sera, revealed a strong reactivity of the 16-17kD components, together with other minor bands at lower MW. Further analysis of the N-terminal aminoacid sequence would provide a full identification of these native allergens. [ABSTRACT FROM AUTHOR]
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Database:	MedicLatina

Description
Abstract:	The domestic mite Blomia tropicalis (Bt) is an important source of allergens causing allergic diseases in tropical countries. More than 20 allergens have been detected in the extract of Bt. By using the rDNA technology, 3 allergens with similar Molecular Weight around 14kD have been obtained and characterized; one of them (Blo-t-5) is thought to be a major allergen. Nevertheless, the native analogs of these recombinant proteins have not been isolated nor identified. The aim of the present work was to isolate the native low molecular allergens (14-17kD) of Bt and to characterize their IgE-binding activity. A partially purified preparation (LMWF) was obtained by precipitation of the freeze-dried allergen extract in 50% ammonium sulfate, and subsequent gelfiltration of the supernatant in Sephadex-G50. The analysis by SDS-PAGE in non-reducing conditions revealed a major 14kD band, which was resolved in reducing conditions, in two close bands at 16-17kD. This band accounted for 80% of the total protein content. The UV absorbance spectrum peaked at 214 and 256nm. The LMWF was able to inhibit up to 69% of total IgE-binding activity of the extract, as measured by IgE-inhibition ELISA, using a serum pool of allergic patients to Bt. Up to 60% IgE reactivity was found by ELISA, to tested sera from 42 allergic subjects. The Western Blotting analysis of selected sera, revealed a strong reactivity of the 16-17kD components, together with other minor bands at lower MW. Further analysis of the N-terminal aminoacid sequence would provide a full identification of these native allergens. [ABSTRACT FROM AUTHOR]
ISSN:	1025028X