ALTEX :: Alternatives to Animal Experiments

1996, Supplement

Production and Purification of Mouse IgG Subclass Specific Chicken Egg Yolk Antibodies Using Indirect Affinity Chromatography Method with Protein G Sepharose

Michael H. Erhard¹, Peter Schmidt², Andrea Hofmann¹, Manfred Stangassinger¹, and Uli Lösch¹

¹Institute of Veterinary Physiology; ²Institute of Veterinary Pathology, Ludwig-Maximilians-University, D-Munich

SUMMARY

Chicken antibodies have been reported to be an excellent alternative to the mammalian antibodies, especially if the antigen is from mammalians. Because chicken immunoglobulin Y (IgY) does not bind to protein G, in this study a indirect method for the purification of murine immunoglobulin G (IgG) subclass specific egg yolk antibodies is described. After preincubation of the chicken egg antibodies (e.g. from hens immunized against murine IgG1) with the non corresponding murine IgG subclasses (e.g. IgG2a and IgG2b) and purification with protein G sepharose, subclass specific chicken antibodies (e.g. against IgG1) could be isolated from precipitated egg yolk antibodies. Specificities of the egg yolk antibodies were determined in a competitive ELISA and immunoblot. Therefore, the indirect affinity chromatography with protein G provides a efficient method for purifying specific chicken egg antibodies against mammalian IgG subclasses. (article in english)

Keywords: affinity chromatograhy, antibody purification, egg yolk antibody, IgY, murine IgG subclasses, protein G