Abstract
We have produced the recombinant intracellular domain of human IA-2 (IA-2ic), a diabetes-associated autoantigen, in plants. This was achieved by transient expression using agroinfiltration of Nicotiana benthamiana plants. The resulting plant-derived IA-2ic had the expected size, reacted with polyclonal and monoclonal antibodies specific to human IA-2ic and competitively inhibited radiolabeled IA-2ic in an immunoprecipitation assay. The expression level of recombinant IA-2ic was estimated to be 0.5% of the total soluble protein (TSP). Transient expression in plants has the potential to produce a large amount of human IA-2ic protein at low cost in a short period of time.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Antibodies, Monoclonal
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Autoantibodies
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Autoantigens / chemistry
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Autoantigens / genetics*
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Autoantigens / immunology*
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Base Sequence
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DNA Primers / genetics
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Diabetes Mellitus, Type 1 / immunology
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Gene Expression
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Humans
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In Vitro Techniques
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Membrane Proteins / chemistry
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Membrane Proteins / genetics*
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Membrane Proteins / immunology*
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Nicotiana / genetics*
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Plants, Genetically Modified
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Protein Engineering
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Protein Structure, Tertiary
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Protein Tyrosine Phosphatase, Non-Receptor Type 1
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Protein Tyrosine Phosphatases / chemistry
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Protein Tyrosine Phosphatases / genetics*
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Protein Tyrosine Phosphatases / immunology*
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Receptor-Like Protein Tyrosine Phosphatases, Class 8
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Recombinant Proteins / chemistry
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Recombinant Proteins / genetics
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Recombinant Proteins / immunology
Substances
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Antibodies, Monoclonal
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Autoantibodies
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Autoantigens
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DNA Primers
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ICA512 autoantibody
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Membrane Proteins
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Recombinant Proteins
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PTPRN protein, human
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Protein Tyrosine Phosphatase, Non-Receptor Type 1
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Protein Tyrosine Phosphatases
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Receptor-Like Protein Tyrosine Phosphatases, Class 8