Production and purification of mouse IgG subclass specific chicken egg yolk antibodies using a new indirect affinity chromatography method with protein G

Abstract

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 immunised against murine IgG1) with the non corresponding murine IgG subclasses (e.g. against 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 an efficient method for purifying specific chicken egg antibodies against mammalian IgG subclasses.