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

摘要

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 (R) 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. (C) 2009 Elsevier B.V. All rights reserved.