Abstract
Single-chain antibodies consist of the variable, antigen-binding domains of antibodies joined to a continuous polypeptide by genetically engineered peptide linkers. We have used the flexible interdomain linker region of a fungal cellulase to link together the variable domains of an anti-2-phenyloxazolone IgG1 and show here that the resulting single-chain antibody is efficiently secreted and released to the culture medium of Escherichia coli. The yield of affinity-purified single-chain antibody is 1-2 mg/l of culture medium and its affinity and stability are comparable to those of the corresponding native IgG.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Antibody Specificity
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Base Sequence
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Cellulose 1,4-beta-Cellobiosidase
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Enzyme-Linked Immunosorbent Assay
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Escherichia coli / genetics*
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Glycoside Hydrolases / genetics*
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Hydrogen-Ion Concentration
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Immunoglobulin G / genetics*
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Immunoglobulin G / immunology
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Molecular Sequence Data
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Oxazolone / analogs & derivatives
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Oxazolone / immunology
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Protein Conformation
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Protein Denaturation
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Recombinant Fusion Proteins* / isolation & purification
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Trichoderma / enzymology
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Trichoderma / genetics*
Substances
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Immunoglobulin G
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Recombinant Fusion Proteins
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2-phenyloxazolone
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Oxazolone
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Glycoside Hydrolases
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Cellulose 1,4-beta-Cellobiosidase