title: Antibody capture by mixed-mode chromatography: a comprehensive study from determination of optimal purification conditions to identification of contaminating host cell proteins.
abstract: We evaluated mixed mode chromatography for the capture of recombinant antibodies from CHO cell culture supernatants. We studied PPA HyperCel, HEA HyperCel, MEP HyperCel and Capto adhere resins, which all contain hydrophobic and cationic groups. A microplate approach combined with DoE modeling allowed the exploration of the complex behaviors of these mixed mode resins. Optimal conditions for antibody purification and host cell proteins (HCPs) elimination were determined and then directly up-scaled to laboratory columns. Then we used mass spectrometry to identify the major HCPs potentially coeluted with the antibody. Differences between the four resins in terms of amount, complexity and identity of the HCPs present in the elution fractions were investigated.
authors: Université de Bordeaux, EA 4135, Bordeaux F-33000, France.
authors: Jerome
authors: J
authors: Pezzini
authors: Gilles
authors: G
authors: Joucla
authors: René
authors: R
authors: Gantier
authors: Magali
authors: M
authors: Toueille
authors: Anne-Marie
authors: AM
authors: Lomenech
authors: Caroline
authors: C
authors: Le Sénéchal
authors: Bertrand
authors: B
authors: Garbay
authors: Xavier
authors: X
authors: Santarelli
authors: Charlotte
authors: C
authors: Cabanne
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000302:isolation & purification*; D016466:CHO Cells; D002853:Chromatography, Liquid / Q000295:instrumentation / Q000379:methods; D006224:Cricetinae; D003412:Cricetulus; D004553:Electric Conductivity; D057166:High-Throughput Screening Assays; D006863:Hydrogen-Ion Concentration; D057927:Hydrophobic and Hydrophilic Interactions; D019995:Laboratory Chemicals; D013058:Mass Spectrometry; D008956:Models, Chemical; D011506:Proteins / Q000737:chemistry / Q000145:classification*; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification*; D015203:Reproducibility of Results
chemical_list: D000911:Antibodies, Monoclonal; D019995:Laboratory Chemicals; D011506:Proteins; D011994:Recombinant Proteins