PROCÉDÉ DE FABRICATION DE DIMÈRES ET DE MULTIMÈRES PAR AUGMENTATION DE LA PRODUCTION DE PONTS DE LIAISON DANS UN COMPLEXE DE MONOMÈRES MULTIPLES ET DE CHAÎNES RÉCURRENTES D'UN ADHÉSIF D'UN TYPE ADHÉRANT SPÉCIFIQUEMENT À DES MONOMÈRES

摘要

The present invention relates to a method for manufacturing multimers by making repeat chains comprising repeatedly linked affinity domains binding specifically to monomers, and by using the same to create a repeat chain/multiple-monomer complex created from the repeat chains and a multiple number of monomers, thereby facilitating the formation of bond bridges between the monomers in the complex to produce inter-monomeric bond bridged multimer. The present invention relates to a method for manufacturing multimers by making repeat chain recombinant proteins resulting from the repeated linking affinity domain proteins binding specifically to protein monomers, and by using the same to create a repeat chain/multiple-monomer complex created from the repeat chains and a multiple number of monomers, thereby facilitating the formation of bond bridges between the monomers in the complex to produce inter-monomeric bond bridged multimer. More specifically, the present invention relates to a method for manufacturing multimers by making a repeat chain recombinant proteins resulting from the repeated linking affinity protein domains binding specifically to protein monomers, and by using the same to create a repeat chain/multiple-monomer complex created from the repeat chains and a multiple number of monomers, thereby facilitating the formation of disulphide bond bridges between the monomers in the complex to produce inter-monomeric disulphide bond bridged multimer. More specifically, it relates to a method for manufacturing dimers and multimers by creating repeat chain recombinant proteins resulting from the repeated linking of the domain III (the Fab binding domain) of Streptococcal protein G, and by using the same as binding matrix (scaffold, skeleton) to create a repeat chain/multiple-monomer complex created from the repeat chains and a multiple number of monomers derived from antibody, thereby facilitating the formation of disulphide bond bridges between the monomers in the complex to produce inter-monomeric disulphide bond bridged multimer. The method of the present invention can be used to advantage in the large-volume manufacture of disulphide bond bridge dimers since it gives up to a 200 fold improvement in the yield of disulphide bond bridge dimers as compared with known refolding methods of the prior art.