Chicken Antibodies: a useful tool for antigen capture ELISA to detect bovine leukaemia virus without cross-reaction with other mammalian antibodies

M. JULIARENA, S. GUTIERREZ AND C. CERIANI

VETERINARY RESEARCH COMMUNICATIONS

Springer Netherlands

Año: 2007 vol. 31 p. 43 - 51

0165-7380

The 24 kDa protein from the gag of the bovine leukaemia virus was cloned and expressed as a fusion protein GST-p24. This recombinant protein was then used to immunize a Leghorn chicken. The partially purified chicken anti-GST IgY was used to develop a solid-phase assay by binding the IgY to an ELISA plate. When the fusion protein contacts the antibody, it binds it by its N-terminal, leaving the C-terminal, which carries the sequence that acts as a capture antigen in solution maximally exposed, reducing the risk of epitope masking. The conditions of the fusion protein on the solid phase maximize the presentation of the antigens´ epitopes in solution. For the first time, a system has been developed with a non-mammalian coating antibody. Besides optimizing the recognition of low-molecular-weight antigens synthesized as fusion proteins, it avoids cross-reactions with commonly used secondary antibodies, mostly raised in mammalian hosts.