Method for manufacturing dimers and multimers in complexes of multiple monomers and repeat chains of affinity domains binding specifically to monomers with increasing the formation of inter-monomeric bond bridges among monomers

摘要

The present invention relates to a method for manufacturing multimers in which a plurality of monomers are linked by bond bridges, by making repeating chains comprising repeatedly linking adherends of a type specifically adhering to monomers, and by using the same to create a repeating chain/multiple-monomer complex created from the repeating chains and a number of monomers, thereby facilitating the formation of bond bridges between the monomers in the complex. More specifically, the present invention relates to a method for manufacturing multimers in which a plurality of monomers are linked by bond bridges, by making a repeating-chain recombinant protein resulting from the repeated linking of an adherent protein of a type specifically adhering to protein monomers, and by using the same to create a repeating chain/multiple-monomer complex created from the repeating chains and a number of monomers, thereby facilitating the formation of bond bridges between the monomers in the complex. More specifically, the present invention relates to a method for manufacturing multimers in which a plurality of monomers are linked by disulphide bond bridges, by making a repeating-chain recombinant protein resulting from the repeated linking of an adherent protein of a type specifically adhering to protein monomers, and by using the same to create a repeating chain/multiple-monomer complex created from the repeating chains and a number of monomers, thereby facilitating the formation of disulphide bond bridges between the monomers in the complex. More specifically, it relates to a method for manufacturing dimers and multimers in which a plurality of monomers are linked by disulphide bond bridges, by creating repeating chains, and multi-monomer complexes thereof, from monomers of an antibody protein through the use of a skeleton comprising a recombinant repeating chain protein resulting from the repeated linking of the domain III (the Fab adhering domain) of Streptococcal protein G, thereby facilitating the formation of disulphide bond bridges between the monomers in the complex. 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.