Cloning and expression of recombinant soluble proteins could be quite a difficult task, especially when it comes to reliably detect minute amounts of the soluble protein in the supernatant of transfected mammalian cells. Timing and sensitivity are of the essence in order to optimise the benefits/costs balance and to decide which clones to grow further and which ones to discard. Here we propose a modified inhibition assay. The key feature of this approach is the development of a sensitive and quantitative test to detect the presence of the recombinant soluble protein by exploiting its ability to compete with the binding of a specific monoclonal antibody to a target cell. The described procedure is a sensitive, efficient, dependable and low cost method.
机构:
Manchester Inst Biotechnol, Fac Life Sci, Prot Express Facil, 131 Princess St, Manchester M1 7DN, Lancs, EnglandManchester Inst Biotechnol, Fac Life Sci, Prot Express Facil, 131 Princess St, Manchester M1 7DN, Lancs, England
McKenzie, Edward A.
Abbott, W. Mark
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Peak Prot Ltd, Alderley Pk, Macclesfield SK10 4TG, Cheshire, EnglandManchester Inst Biotechnol, Fac Life Sci, Prot Express Facil, 131 Princess St, Manchester M1 7DN, Lancs, England
机构:
Xinxiang Med Univ, Sch Life Sci & Technol, Xinxiang, Peoples R China
Int Joint Res Lab Recombinant Pharmaceut Prot Expr, Xinxiang, Peoples R ChinaXinxiang Med Univ, Sch Life Sci & Technol, Xinxiang, Peoples R China
Wei, Mian
Mi, Chun-Liu
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Int Joint Res Lab Recombinant Pharmaceut Prot Expr, Xinxiang, Peoples R ChinaXinxiang Med Univ, Sch Life Sci & Technol, Xinxiang, Peoples R China
Mi, Chun-Liu
Jing, Chang-Qin
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Xinxiang Med Univ, Sch Life Sci & Technol, Xinxiang, Peoples R ChinaXinxiang Med Univ, Sch Life Sci & Technol, Xinxiang, Peoples R China
Jing, Chang-Qin
Wang, Tian-Yun
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Int Joint Res Lab Recombinant Pharmaceut Prot Expr, Xinxiang, Peoples R ChinaXinxiang Med Univ, Sch Life Sci & Technol, Xinxiang, Peoples R China
机构:
Univ Oxford, Div Struct Biol, Wellcome Ctr Human Genet, Oxford, England
Univ Bordeaux, Interdisciplinary Inst Neurosci, Bordeaux, France
CNRS, UMR 5297, Interdisciplinary Inst Neurosci, Bordeaux, FranceUniv Oxford, Div Struct Biol, Wellcome Ctr Human Genet, Oxford, England
Elegheert, Jonathan
Behiels, Ester
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Univ Oxford, Div Struct Biol, Wellcome Ctr Human Genet, Oxford, England
Univ Bordeaux, Interdisciplinary Inst Neurosci, Bordeaux, France
CNRS, UMR 5297, Interdisciplinary Inst Neurosci, Bordeaux, FranceUniv Oxford, Div Struct Biol, Wellcome Ctr Human Genet, Oxford, England
Behiels, Ester
Bishop, Benjamin
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Univ Oxford, Div Struct Biol, Wellcome Ctr Human Genet, Oxford, EnglandUniv Oxford, Div Struct Biol, Wellcome Ctr Human Genet, Oxford, England
Bishop, Benjamin
Scott, Suzanne
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Univ Oxford, Div Struct Biol, Wellcome Ctr Human Genet, Oxford, England
MRC, Mol Biol Lab, Cambridge Biomed Campus, Cambridge, EnglandUniv Oxford, Div Struct Biol, Wellcome Ctr Human Genet, Oxford, England
Scott, Suzanne
Woolley, Rachel E.
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Univ Oxford, Div Struct Biol, Wellcome Ctr Human Genet, Oxford, EnglandUniv Oxford, Div Struct Biol, Wellcome Ctr Human Genet, Oxford, England
Woolley, Rachel E.
Griffiths, Samuel C.
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Univ Oxford, Div Struct Biol, Wellcome Ctr Human Genet, Oxford, EnglandUniv Oxford, Div Struct Biol, Wellcome Ctr Human Genet, Oxford, England
Griffiths, Samuel C.
Byrne, Eamon F. X.
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Univ Oxford, Div Struct Biol, Wellcome Ctr Human Genet, Oxford, England
Stanford Univ, Dept Bioengn, Stanford, CA 94305 USAUniv Oxford, Div Struct Biol, Wellcome Ctr Human Genet, Oxford, England
Byrne, Eamon F. X.
Chang, Veronica T.
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MRC, Mol Biol Lab, Cambridge Biomed Campus, Cambridge, EnglandUniv Oxford, Div Struct Biol, Wellcome Ctr Human Genet, Oxford, England
Chang, Veronica T.
Stuart, David I.
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Univ Oxford, Div Struct Biol, Wellcome Ctr Human Genet, Oxford, EnglandUniv Oxford, Div Struct Biol, Wellcome Ctr Human Genet, Oxford, England
Stuart, David I.
Jones, E. Yvonne
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Univ Oxford, Div Struct Biol, Wellcome Ctr Human Genet, Oxford, EnglandUniv Oxford, Div Struct Biol, Wellcome Ctr Human Genet, Oxford, England
Jones, E. Yvonne
Siebold, Christian
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Univ Oxford, Div Struct Biol, Wellcome Ctr Human Genet, Oxford, EnglandUniv Oxford, Div Struct Biol, Wellcome Ctr Human Genet, Oxford, England
Siebold, Christian
Aricescu, A. Radu
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Univ Oxford, Div Struct Biol, Wellcome Ctr Human Genet, Oxford, England
MRC, Mol Biol Lab, Cambridge Biomed Campus, Cambridge, EnglandUniv Oxford, Div Struct Biol, Wellcome Ctr Human Genet, Oxford, England