Abstract
In vitro display technology is a powerful tool for discovery and optimisation of novel antibodies. With increasing demands on various binding molecules in proteomics studies, techniques for a large-scale generation of antibodies or antibody fragments are needed. Here, we describe a novel method for parallel generation of different antibody fragments (scFv) by integrating cell-free ribosome display with array technology. We have demonstrated the procedure by successfully isolating scFv antibodies specific to 16 different cancer biomarkers via a single process. Our results provide proof of principle for multiple production of various scFv antibodies simultaneously.
Keywords:
Cell free ribosome display; Dot blot screening; Multiple antibody generation in vitro; Protein arrays.
© 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
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 / immunology
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Antigens / genetics
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Antigens / immunology
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Antigens / metabolism*
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Biomarkers, Tumor / genetics
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Biomarkers, Tumor / immunology
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Biomarkers, Tumor / metabolism
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Blotting, Western
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Cell-Free System
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Enzyme-Linked Immunosorbent Assay
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Escherichia coli / genetics
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Humans
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Neoplasms / genetics
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Neoplasms / immunology
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Neoplasms / metabolism*
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Peptide Library
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Protein Array Analysis / methods*
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Protein Binding / immunology
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Protein Biosynthesis
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Recombinant Proteins / immunology
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Recombinant Proteins / metabolism
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Reproducibility of Results
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Ribosomes / genetics
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Ribosomes / metabolism*
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Sequence Homology, Amino Acid
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Single-Chain Antibodies / genetics
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Single-Chain Antibodies / immunology
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Single-Chain Antibodies / metabolism*
Substances
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Antigens
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Biomarkers, Tumor
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Peptide Library
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Recombinant Proteins
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Single-Chain Antibodies