Abstract
A bispecific disulfide-stabilized Fv antibody fragment (dsFv-dsFv') consisting of two different disulfide-stabilized Fv antibody fragments connected by flexible linker peptides was produced by secretion of three polypeptide chains into the periplasm of Escherichia coli. The dsFv-dsFv' molecules were enriched by immobilized metal affinity chromatography and further purified by anion-exchange chromatography. The recombinant antibody constructs retained the two parental antigen binding specificities and were able to cross-link the two different antigens. The described dsFv-dsFv' design might be of particular value for therapeutic in vivo applications since improved stability is expected to be combined with minimal immunogenicity.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Antibodies, Bispecific / biosynthesis
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Antibodies, Bispecific / chemistry*
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Antibodies, Bispecific / isolation & purification
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Antibody Affinity
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Disulfides / chemistry
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Enzyme-Linked Immunosorbent Assay
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Escherichia coli / immunology*
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Genetic Vectors / chemical synthesis
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Immunoblotting
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Immunoglobulin Variable Region / biosynthesis
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Immunoglobulin Variable Region / chemistry*
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Immunoglobulin Variable Region / isolation & purification
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Mice
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Mutagenesis, Insertional
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Peptides / chemistry
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Recombinant Proteins / biosynthesis
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Recombinant Proteins / chemistry
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Recombinant Proteins / isolation & purification
Substances
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Antibodies, Bispecific
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Disulfides
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Immunoglobulin Variable Region
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Peptides
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Recombinant Proteins