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, 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.
Pezzini
J
J
EA4135, ESTBB, Université de Bordeaux, 146 rue Léo Saignat, 33076 Bordeaux Cedex, France.
Brenac Brochier
V
V
Barrouillet
M P
MP
Cerruti
M
M
Clofent-Sanchez
G
G
Schapman
A
A
Topol
A
A
Robert
R
R
Cabanne
C
C
Cerruti
P
P
Santarelli
X
X
D000818:Animals; D016367:Baculoviridae / Q000235:genetics* / Q000378:metabolism; D002460:Cell Line; D002853:Chromatography, Liquid / Q000295:instrumentation / Q000379:methods*; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012117:Resins, Synthetic / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D018411:Spodoptera
D007140:Immunoglobulin Fab Fragments; D011993:Recombinant Fusion Proteins; D012117:Resins, Synthetic