caa a22 4500 463236947 CHVBK 20180405153300.0 cr unu---uuuuu 170326e20070101xx s 000 0 eng 10.1007/s11259-006-3422-1 doi (NATIONALLICENCE)springer-10.1007/s11259-006-3422-1 Chicken Antibodies: A Useful Tool for Antigen Capture ELISA to Detect Bovine Leukaemia Virus without Cross-reaction with Other Mammalian Antibodies [Elektronische Daten] [M. Juliarena, S. Gutierrez, C. Ceriani] 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. Springer Science + Business Media, Inc., 2007 BLV nationallicence chicken IgY nationallicence ELISA nationallicence glutathione S -transferase nationallicence pGEX recombinant peptides nationallicence BLV : bovine leukaemia virus nationallicence GST : glutathione S-transferase nationallicence PCR : polymerase chain reaction nationallicence FLK : fetal lamb kidney nationallicence GS4B : glutathione agarose 4B nationallicence IPTG : isopropyl-β -d-thiogalactopyranoside nationallicence PMSF : phenylmethylsulphonyl fluoride nationallicence FCA : Freund's complete adyuvant nationallicence BCI/TNBT : 5-bromo-4-chloro-3-indolyl phosphate/tetranitroblue tetrazolium nationallicence SDS-PAGE : sodium dodecyl sulphate-polyacrylamide gel electrophoresis nationallicence TMB : 3,3′,5,5′-tetramethylbenzidine nationallicence Juliarena M. Area Virologia, Dpto. SAMP, Tandil, Argentina aut Gutierrez S. Area Virologia, Dpto. SAMP, Tandil, Argentina aut Ceriani C. Dpto. Ciencias Biológicas, Facultad de Ciencias Veterinarias UNCPBA, Tandil, Argentina aut Veterinary Research Communications Kluwer Academic Publishers 31/1(2007-01-01), 43-51 0165-7380 31:1<43 2007 31 11259 https://doi.org/10.1007/s11259-006-3422-1 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11259-006-3422-1 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Juliarena M. Area Virologia, Dpto. SAMP, Tandil, Argentina aut NATIONALLICENCE 700 1- Gutierrez S. Area Virologia, Dpto. SAMP, Tandil, Argentina aut NATIONALLICENCE 700 1- Ceriani C. Dpto. Ciencias Biológicas, Facultad de Ciencias Veterinarias UNCPBA, Tandil, Argentina aut NATIONALLICENCE 773 0- Veterinary Research Communications Kluwer Academic Publishers 31/1(2007-01-01), 43-51 0165-7380 31:1<43 2007 31 11259 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer