Servizio Comunitario di Informazione in materia di Ricerca e Sviluppo - CORDIS

Recovery of recombinant antibody produced in crop plants

The project involves the provision of generic processing technologies for affinity-based separation of antibody fragments produced in plants, by exploiting small stable peptides which resemble the natural epitope recognized by the antibody. The peptides are selected on the basis that they are inexpensive to produce in bulk, and designed with the requirements of being linked with large scale supports to allow quick, reliable recovery and also multiple release of the affinity medium. Monoclonal antibodies against the model antigens glucose oxidase (GOx) and follicle stimulating hormone (FSH) have been produced and evaluated for peptide ligands by both Pepscan assay and phage displayed random peptide libraries. From linear epitope sequences for GOx and FSH antibodies, appropriate peptides have been synthesized and checked for reactions. Immunoassays appropriate to the prospective affinity processes are being designed for purification technology. An issue has arisen as to whether peptides should be linear or cyclized for most appropriate binding reaction affinity. Combinatorial phage rabbit and chicken antibody libraries are being established from immunocompetent mice to select phages producing antibody fragments that react with GOx/FSH peptide ligands selected. Studies are also ongoing with rabbits and chickens to examine their relevance as sources of antibody genes. Vectors for cloning and expression of scFvs in the yeast Pichia pastoris and plants have been prepared for future use in optimizing heterologous expression of suitable scFvs. Fermenters have been commissioned and production of mouse scFv antibody fragments successfully achieved in quantities that will facilitate the development of affinity processing media.


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