Expression of a bispecific dsFv-dsFv' antibody fragment in Escherichia coli |
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Authors: | Schmiedl Andreas; Breitling Frank; Dubel Stefan |
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Affiliation: | 1 Universität Heidelberg, Institut für Molekulare Genetik, Im Neuenheimer Feld 230, 69120 Heidelberg and
3 German Cancer Research Center, Im Neuenheimer Feld 280, 69120 Heidelberg, Germany |
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Abstract: | A bispecific disulfide-stabilized Fv antibody fragment (dsFvdsFv')consisting of two different disulfide-stabilized Fv antibodyfragments connected by flexible linker peptides was producedby secretion of three polypeptide chains into the periplasmof Escherichia coli. The dsFvdsFv' molecules were enrichedby immobilized metal affinity chromatography and further purifiedby anion-exchange chromatography. The recombinant antibody constructsretained the two parental antigen binding specificities andwere able to cross-link the two different antigens. The describeddsFvdsFv' design might be of particular value for therapeuticin vivo applications since improved stability is expected tobe combined with minimal immunogenicity. |
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