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(The FASEB Journal. 1999;13:81-88.)
© 1999 FASEB

Research Communications

General applicability of chicken egg yolk antibodies: the performance of IgY immunoglobulins raised against the hypoxia-inducible factor 1{alpha}

Gieri Camenischa, Mauro Tinia, Dmitri Chilova, Ivica Kvietikovaa, Vickram Srinivasb, Jaime Carob, Patrick Spielmanna, Roland H. Wengera and Max Gassmanna,1

a Institute of Physiology, University of Zürich-Irchel, CH-8057 Zürich, Switzerland
b Department of Medicine, Thomas Jefferson University, Philadelphia, Pennsylvania 19107–5099, USA

Avian embryos and neonates acquire passive immunity by transferring maternal immunoglobulins from serum to egg yolk. Despite being a convenient source of antibodies, egg yolk immunoglobulins (IgY) from immunized hens have so far received scant attention in research. Here we report the generation and rapid isolation of IgY from the egg yolk of hens immunized against the {alpha} subunit of the human hypoxia-inducible factor 1 (HIF-1{alpha}). Anti-HIF-1{alpha} IgY antibodies were affinity purified and tested for their performance in various applications. Abundant HIF-1{alpha} protein was detected by Western blot analysis in nuclear extracts derived from hypoxic cells of human, mouse, monkey, swine, and dog origin whereas in hypoxic quail and frog cells, the HIF-1{alpha} signal was weak or absent, respectively. In electrophoretic mobility shift assays, affinity-purified IgY antibody was shown to recognize the native HIF-1 (but not the related HIF-2) complex that specifically binds an oligonucleotide containing the HIF-1 DNA binding site. Furthermore, IgY antibody immunoprecipitated HIF-1{alpha} from hypoxic cell extracts. Immunofluorescence experiments using IgY antibody allowed the detection of HIF-1{alpha} in the nucleus of hypoxic COS-7 cells. For comparison, the application of a mouse monoclonal antibody raised against the identical HIF-1{alpha} fragment was more restricted. Because chicken housing is inexpensive, egg collection is noninvasive, isolation and affinity purification of IgY antibodies are fast and simple, and the applicability of IgY is widespread, immunization of hens represents an excellent alternative for the generation of polyclonal antibodies.—Camenisch, G., Tini, M., Chilov, D., Kvietikova, I., Srinivas, V., Caro, J., Spielmann, P., Wenger, R. H., Gassmann, M. General applicability of chicken egg yolk antibodies: the performance of IgY immunoglobulins raised against the hypoxia-inducible factor 1{alpha}. FASEB J. 13, 81–88 (1999)


Key Words: monoclonal antibody • affinity purification • Western blotting • EMSA • immunoprecipitation • immunofluorescence




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