MARIA, Sophie; JOUCLA, Gilles; GARBAY, Bertrand; DIERYCK, Wilfrid; LOMENECH, Anne Marie; SANTARELLI, Xavier; CABANNE, Charlotte

doi:10.1016/j.chroma.2015.03.018

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MARIA, Sophie
Biotechnologie des protéines recombinantes à visée santé

JOUCLA, Gilles

GARBAY, Bertrand

Biotechnologie des protéines recombinantes à visée santé
Laboratoire de Chimie des Polymères Organiques [LCPO]
Team 3 LCPO : Polymer Self-Assembly & Life Sciences

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Journal of Chromatography A. 2015, vol. 1393, p. 57-64

Elsevier

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An innovative process to purify mAb from CHO cell culture supernatant was developed. This three-step process involved two mixed mode resins and an anion exchange membrane. We used a human IgG mixture to determine the optimal conditions for each purification step. Thereafter, the whole process was evaluated and improved for the purification of a recombinant mAb produced in the supernatant of CHO cells. Once optimized, yield and purity of 88% and 99.9%, respectively were comparable to those obtained in a conventional process based on a capture step using protein A. In addition, aggregates, HCPs and DNA levels in the purified fraction were below regulatory specifications. Then we used mass spectrometry to identify contaminating proteins in the antibody fraction in order to highlight the behavior of HCPs. (C) 2015 Elsevier B.V. All rights reserved.< Leer menos

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ADSORBENTS

IDENTIFICATION

AGGREGATION

Mixed mode chromatography

Antibody

Host cell proteins

Purification process

CHARGE INDUCTION CHROMATOGRAPHY

HOST-CELL PROTEINS

SEPARATION

CAPTURE

BINDING

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Purification process of recombinant monoclonal antibodies with mixed mode chromatography

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