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Production of an egg yolk antibody against Parietaria judaica 2 allergen*

Posted by on in 2009
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 R. Alessandro*,1, A. Gallo*, M. Barranca*, S. Principe*, S. Taverna*, G. Duro{dagger}, G. Cassata{ddagger}, M. Becchi§, S. Fontana* and G. De Leo* Poult Sci. 2009 88:1773-1778.

* Università di Palermo, Sezione di Biologia e Genetica, Dipartimento di Biopatologia e Metodologie Biomediche, 90133 Palermo, Italy; {dagger} Istituto di Biomedicina ed Immunologia Molecolare "Alberto Monroy," Consiglio Nazionale delle Ricerche, 90146 Palermo, Italy; {ddagger} Istituto Zooprofilattico Sperimentale della Sicilia "A. Mirri,"90129 Palermo, Italy; and § UMR 5086 CNRS, Institut de Biologie et Chimie des Proteines, IFR128, Lyon, France

1 Corresponding author: ricale@unipa.it

Abstract

Specific antibodies are essential tools for studying proteins as well as for diagnostic research in biomedicine. The egg yolk of immunized chicken is an inexpensive source of high-quality polyclonal antibodies. The 12-kDa Parietaria judaica 2 allergen was expressed as a fusion protein and was used to immunize Leghorn chickens. In this paper, we show, using 2-dimensional gel electrophoresis and immunoblotting, that chicken antibodies raised against a recombinant allergen can be used to recognize similar proteins from a pollen raw extract. Allergen identity was confirmed by nanoLC-nanospray-tandem mass spectrometry analysis. Our data demonstrate for the first time that a synergistic combination of molecular biology, 2-dimensional PAGE, and use of nonmammalian antibodies represents a powerful tool for reliable identification of allergens.

Key Words: antibody • allergy • Parietaria judaica • pollen

*Note: The chicken IgY used in this publication was purified using the Eggspress IgY purification kit manufactured by Gallus Immunotech Inc.

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