Abstract
The development of a capture step of a human recombinant F(ab')(2) produced and expressed in baculovirus-infected cells was investigated by screening three mixed-mode chromatography sorbents (HEA HyperCel, PPA HyperCel and MEP HyperCel) and two ion exchangers (Q Ceramic HyperD F, S Ceramic HyperD F) sorbents using a 96-well plate format and SELDI-MS. HEA HyperCel gave the best separation performance therefore the conditions tested in micro-plate were transferred to laboratory scale chromatographic experiments, confirming that the recombinant F(ab')(2) was effectively captured on the mixed-mode sorbent without any pre-treatment of the crude extract with a 82% recovery and a 39-fold purification.
Publication types
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Evaluation Study
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Baculoviridae / genetics*
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Baculoviridae / metabolism
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Cell Line
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Chromatography, Liquid / instrumentation
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Chromatography, Liquid / methods*
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Gene Expression
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Genetic Vectors / genetics
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Genetic Vectors / metabolism
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Humans
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Immunoglobulin Fab Fragments / chemistry
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Immunoglobulin Fab Fragments / genetics
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Immunoglobulin Fab Fragments / isolation & purification*
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Immunoglobulin Fab Fragments / metabolism
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Protein Binding
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Recombinant Fusion Proteins / chemistry
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / isolation & purification
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Recombinant Fusion Proteins / metabolism
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Resins, Synthetic / chemistry*
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Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods*
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Spodoptera
Substances
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Immunoglobulin Fab Fragments
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Recombinant Fusion Proteins
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Resins, Synthetic