Production of an egg yolk antibody against Parietaria judaica 2 allergen.
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.
Alessandro
R
R
Università di Palermo, Sezione di Biologia e Genetica, Dipartimento di Biopatologia e Metodologie Biomediche, Palermo, Italy. ricale@unipa.it
Gallo
A
A
Barranca
M
M
Principe
S
S
Taverna
S
S
Duro
G
G
Cassata
G
G
Becchi
M
M
Fontana
S
S
De Leo
G
G
D000818:Animals; D000906:Antibodies / Q000276:immunology*; D052179:Antigens, Plant / Q000276:immunology*; D002645:Chickens / Q000276:immunology* / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004530:Egg Yolk / Q000378:metabolism*; D005260:Female; D007136:Immunoglobulins / Q000276:immunology* / Q000378:metabolism; D032403:Parietaria / Q000737:chemistry* / Q000276:immunology; D011058:Pollen / Q000276:immunology
D000906:Antibodies; D052179:Antigens, Plant; C016512:IgY; D007136:Immunoglobulins