Abstract
The display of full-length antibody on the cell surface was achieved by fusing a transmembrane domain of the platelet-derived growth factor receptor (PDGFR) to the C-terminus of the heavy chain constant region. We also incorporated a furin cleavage site between the constant region and PDGFR transmembrane domain to obtain secreted antibodies. As a result, antibodies can be expressed simultaneously on the cell surface in a membrane-anchored version for screening and selecting through fluorescence-activated cell sorting (FACS) analysis, as well as in conditioned medium in a secreted version for function analysis.
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 / chemistry
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Antibodies / genetics*
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Antibodies / metabolism
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CHO Cells
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Cell Membrane / metabolism
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Cricetulus
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DNA Restriction Enzymes / genetics
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DNA Restriction Enzymes / metabolism
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Escherichia coli / genetics
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Escherichia coli / metabolism
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Furin / metabolism
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Gene Expression
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Genetic Vectors / chemistry*
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HEK293 Cells
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High-Throughput Screening Assays
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Humans
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Immunoglobulin Heavy Chains / chemistry
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Immunoglobulin Heavy Chains / genetics*
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Immunoglobulin Heavy Chains / metabolism
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Protein Engineering
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Proteolysis
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Receptors, Platelet-Derived Growth Factor / genetics*
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Receptors, Platelet-Derived Growth Factor / metabolism
Substances
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Antibodies
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Immunoglobulin Heavy Chains
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Receptors, Platelet-Derived Growth Factor
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DNA Restriction Enzymes
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FURIN protein, human
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Furin
Grants and funding
This work was partly supported by grants from the National Natural Science Foundation of China (#81173098 to SJ) and from Dgen Biotech Ltd., Hong Kong. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.