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J Chromatogr B Analyt Technol Biomed Life Sci ; 877(24): 2428-34, 2009 Aug 15.
Article in English | MEDLINE | ID: mdl-19467934

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.


Subject(s)
Baculoviridae/genetics , Chromatography, Liquid/methods , Immunoglobulin Fab Fragments/isolation & purification , Resins, Synthetic/chemistry , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods , Animals , Baculoviridae/metabolism , Cell Line , Chromatography, Liquid/instrumentation , Gene Expression , Genetic Vectors/genetics , Genetic Vectors/metabolism , Humans , Immunoglobulin Fab Fragments/chemistry , Immunoglobulin Fab Fragments/genetics , Immunoglobulin Fab Fragments/metabolism , Protein Binding , Recombinant Fusion Proteins/chemistry , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/isolation & purification , Recombinant Fusion Proteins/metabolism , Spodoptera
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