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Lipid Transfer Proteins from Fruit: Cloning, Expression and Quantification.

Authors :
Zuidmeer, Laurian
van Leeuwen, W. Astrid
Budde, Ilona Kleine
Cornelissen, Jessica
Bulder, Ingrid
Rafalska, Ilona
Besolí, Noèlia Telléz
Akkerdaas, Jaap H.
Asero, Riccardo
Rivas, Montserrat Fernandez
Mancebo, Eloina Gonzalez
van Ree, Ronald
Source :
International Archives of Allergy & Immunology. 2005, Vol. 137 Issue 4, p273-281. 9p.
Publication Year :
2005

Abstract

Background: Lipid transfer proteins (LTP) are stable, potentially life-threatening allergens in fruits and many other vegetable foods. The aim of this study was to clone and express recombinant apple LTP (Mal d 3), as has previously been done for peach LTP (Pru p 3) and set up quantitative tests for measuring fruit LTPs. Methods: cDNA for Mal d 3 and Pru p 3 was cloned, expressed in the yeast Pichia pastoris and the resulting proteins were purified via cation exchange chromatography. The immune reactivity of rMal d 3 was compared to nMal d 3 by RAST (inhibition), immunoblotting and basophil histamine release testing. To obtain monoclonal and monospecific polyclonal antibodies, mice and rabbits were immunized with purified nMal d 3. Results: The deduced amino acid sequence of Mal d 3 was identical to the published sequence, Pru p 3 differed at two positions (S9A and S76H). The rMal d 3 had an IgE-binding potency and biological activity close to its natural counterpart. One sandwich ELISA selectively detecting apple LTP and another cross-reactive with cherry, nectarine and hazelnut LTP were developed. In addition, a competitive RIA was developed with polyclonal rabbit antiserum and labeled nMal d 3. Conclusion: rMal d 3 (as shown before for rPru p 3) may be a useful tool for application in component-resolved diagnosis of food allergy. Assays for the measurement of LTP will increase the traceability of this potentially dangerous allergen. Copyright © 2005 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Details

Language :
English
ISSN :
10182438
Volume :
137
Issue :
4
Database :
Academic Search Index
Journal :
International Archives of Allergy & Immunology
Publication Type :
Academic Journal
Accession number :
17934324
Full Text :
https://doi.org/10.1159/000086420