Gene gun-supported DNA immunisation of chicken for straightforward production of poxvirus-specific IgY antibodies

被引:12
作者
Witkowski, Peter T. [1 ]
Bourquain, Daniel R. [1 ]
Hohn, Oliver [2 ]
Schade, Ruediger [3 ]
Nitsche, Andreas [1 ]
机构
[1] Robert Koch Inst, Zentrum Biol Sicherheit & Konsiliarlabor Pockenvi, D-13353 Berlin, Germany
[2] Robert Koch Inst, Zentrum Biol Sicherheit 4, D-13353 Berlin, Germany
[3] Charite, Inst Pharmakol CCM CBF, D-10117 Berlin, Germany
关键词
IgY; Avian antibody; Gene gun; DNA immunisation; Poxvirus; EGG-YOLK ANTIBODIES; UBIQUITIN LIGASE; MOUSE MUSCLE; PLASMID DNA; PROTEIN; VACCINES; MICROPROJECTILES; IMMUNOGENICITY; EXPRESSION; EFFICACY;
D O I
10.1016/j.jim.2008.11.008
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Orthopoxviruses code for numerous immunomodulatory proteins, the structure and function of which are clarified inadequately. Antibodies constitute a potent tool to study such proteins, enabling conclusions on protein location and time course of expression. However, common antibody production in mice or rabbits requires tedious protein expression and injection, as well as blood collection at regular intervals. To simplify this procedure, IgY antibodies specific for poxviral proteins (F1L and p28) were generated by immunisation of chickens, because antibody retrieval from eggs allows the noninvasive generation of huge amounts of antibodies. The main intentions were (i) to decrease invasiveness, (ii) to immunise with native forms of proteins and (iii) to circumvent previous protein expression and purification. Therefore, chicken were immunised with DNA expression vectors coding for conserved domains of the selected proteins delivered for the first time by a gene gun. Four weeks after initial immunisation specific antibodies were found in the egg yolk as proven by immunofluorescence staining of poxvirus-infected cells. The specific IgY titre rose to 1:80,000 and was stable for more than 120 days. With this investigation we present an universal procedure for IgY design and production that can be applied for various issues in the future. (C) 2008 Elsevier B.V. All rights reserved.
引用
收藏
页码:146 / 153
页数:8
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