Immunisation of chicken with the aminoterminal propeptide of bovine procollagen type III. Specificity of of egg yolk antibodies and comparison with immunoassays using rabbit and mouse antibodies

Abstract

Two chicken were immunised with the aminoterminal propeptide of bovine procollagen type III (PIIINP) purified from bovine fetal skin. Both animals developed antibodies binding to either unmodified or iodinated bovine PIIINP, but only one chicken developed antibodies which recognise PIIINP variants in serum. These antibodies were used to establish RIAs to analyse the specificity of these particular antibodies and to compare their specificity with that of published assays, as well as laboratory assay variants utilising different rabbit and mouse antibodies. In comparison to most anti-PIIINP polyclonal antisera from rabbits the chicken antibodies do not bind to the Col-I domain of PIIINP, a characteristic which they share with a monoclonal antibody (MAB 238) produced from a mouse immunised with the same antigen. Like the monoclonal antibody, the chicken antibodies exhibit reactivity against intact serum PIIINP and its high molecular weight variants, most probably pN procollagen type III and procollagen type III. While the monoclonal antibody can only be applied to analyse PIIINP in human sera, the avian antibodies show reactivity to the antigen in both, human and rat sera. The chicken anti PIIINP antibodies described in this study may become a powerful tool to quantitate PIIINP in serum of patients with liver fibrosis and in serum from experimental rat models of fibrogenesis. The fact that both can be analysed with the same assay, gives - for the first time - the opportunity for a direct comparison of the results from human and rat studies designed to evaluate the action of antifibrotic agents.