Rapid screening of purification strategies for the capture of a human recombinant F(ab')(2) expressed in baculovirus-infected cells using a micro-plate approach and SELDI-MS

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[TM], PPA HyperCel and MEP HyperCel) and two ion exchangers (Q Ceramic HyperD F, S Ceramic...

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Veröffentlicht in:Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2009-08, Vol.877 (24), p.2428-2434
Hauptverfasser: Pezzini, J, Brochier, V Brenac, Barrouillet, M P, Cerruti, M, Clofent-Sanchez, G, Schapman, A, Topol, A, Robert, R, Cabanne, C, Cerruti, P, Santarelli, X
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Sprache:eng
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Zusammenfassung: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[TM], 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.
ISSN:1570-0232
DOI:10.1016/j.jchromb.2009.04.034