Abstract
The cDNA of human B lymphocyte stimulator C-terminal peptide (C-BLyS) was amplified by nested PCR from cDNA library of human fetal brain. The expression plasmid pT7450-C-BLyS was constructed and transformed into E. coli BL21 CodonPlus (DE3) RIL which can recognize many rare codons. The C-BLyS protein was expressed as inclusion body in E. coli BL21 CodonPlus (DE3) RIL and the inclusion body of C-BLyS was denatured and then refolded by dialysis and purified by ion exchange chromatography. The refolded and purified C-BLyS can specifically bind with BCMA-Fc, a fusion protein of BLyS receptor and human IgG1 Fc. Furthermore, C-BLyS is proved to be an effective stimulator in mouse splenocytes proliferation in vitro and effective immunostimulant in vivo.
Publication types
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English Abstract
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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B-Cell Activating Factor
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Binding, Competitive
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Cell Division / drug effects
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Chromatography, Ion Exchange
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Dose-Response Relationship, Drug
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Humans
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Immunoglobulin Fc Fragments / immunology
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Immunoglobulin Fc Fragments / metabolism
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Immunoglobulin G / immunology
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Immunoglobulin G / metabolism
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Membrane Proteins / genetics
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Membrane Proteins / immunology*
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Membrane Proteins / pharmacology
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Mice
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Muramidase / immunology
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Muramidase / metabolism
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Protein Folding
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Receptors, Tumor Necrosis Factor / metabolism
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Recombinant Proteins / chemistry
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Recombinant Proteins / isolation & purification*
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Recombinant Proteins / metabolism
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Spleen / cytology
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Spleen / drug effects
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Tumor Necrosis Factor-alpha / genetics
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Tumor Necrosis Factor-alpha / immunology*
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Tumor Necrosis Factor-alpha / pharmacology
Substances
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B-Cell Activating Factor
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BLyS receptor
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Immunoglobulin Fc Fragments
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Immunoglobulin G
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Membrane Proteins
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Receptors, Tumor Necrosis Factor
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Recombinant Proteins
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TNFSF13B protein, human
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Tnfsf13b protein, mouse
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Tumor Necrosis Factor-alpha
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Muramidase