Abstract
Lym-1 single-chain Fv (sFv) can be used for targeted radiodiagnosis and therapy of B-lymphocytic malignancies. Lym-1 sFv was constructed and expressed as a glutathione S-transferase fusion protein, using a (G4S)3 linker connecting the C-terminus of the VH domain and the N-terminus of the VL domain of Lym-1. Six histidine residues and an E Tag epitope were introduced at the C-terminus of the sFv. Lym-1 sFv was purified with glutathione-Sepharose 4B affinity chromatography followed by digestion with thrombin. Lym-1 sFv of 28 kDa was confirmed by Western blotting with anti-(E Tag) monoclonal antibody. An antigen binding assay of Lym-1 and a CD study indicated that it is functionally active.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Amino Acid Sequence
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Antibodies, Monoclonal / chemistry*
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Antibodies, Monoclonal / immunology
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Antigens, Surface / immunology
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B-Lymphocytes / immunology*
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Base Sequence
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Chromatography, Affinity / methods
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Circular Dichroism
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Enzyme-Linked Immunosorbent Assay
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Escherichia coli / metabolism*
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Gene Expression / genetics
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Humans
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Immunoglobulin Variable Region / immunology*
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Leukemia, Lymphoid / immunology
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Leukemia, Lymphoid / therapy
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Molecular Sequence Data
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Protein Structure, Secondary
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Recombinant Proteins / immunology
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Recombinant Proteins / isolation & purification
Substances
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Antibodies, Monoclonal
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Antigens, Surface
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Immunoglobulin Variable Region
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Recombinant Proteins