Abstract
Heterofunctional azide and alkyne PEG-linkers have been synthesized and site specifically conjugated to scFv via a reactive thiol functionality; two scFv were coupled by copper catalyzed 1,3-dipolar cycloaddition to make divalent scFv (di-scFv) with an inter-scFv distance defined to provide divalent binding; antigen binding was maintained for the di-scFv construct and increased several times compared to that of the parent scFv; the cycloaddition reaction reported herein represents an important ligation strategy to covalently link macromolecular proteins and retain sensitive structural conformations.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, U.S. Gov't, Non-P.H.S.
MeSH terms
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Alkynes
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Antigen-Antibody Reactions
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Antigens, Neoplasm / analysis
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Antigens, Neoplasm / immunology
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Antigens, Neoplasm / metabolism
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Azides
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Cell Line, Tumor
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Cross-Linking Reagents / chemistry
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Dimerization
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Enzyme-Linked Immunosorbent Assay
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Female
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Humans
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Immunoglobulin Heavy Chains / chemistry
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Immunoglobulin Light Chains / chemistry
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Immunoglobulin Variable Region / chemistry*
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Immunoglobulin Variable Region / therapeutic use
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Immunohistochemistry
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Male
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Protein Engineering
Substances
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Alkynes
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Antigens, Neoplasm
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Azides
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Cross-Linking Reagents
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Immunoglobulin Heavy Chains
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Immunoglobulin Light Chains
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Immunoglobulin Variable Region