title: Purification process of recombinant monoclonal antibodies with mixed mode chromatography.
abstract: 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.
authors: Univ Bordeaux, BPRVS, EA 4135, F-33000, Bordeaux, France, Bordeaux INP, BPRVS, EA 4135, F-33000, Bordeaux, France.
authors: Sophie
authors: S
authors: Maria
authors: Univ Bordeaux, BPRVS, EA 4135, F-33000, Bordeaux, France, Bordeaux INP, BPRVS, EA 4135, F-33000, Bordeaux, France.
authors: Gilles
authors: G
authors: Joucla
authors: Univ Bordeaux, BPRVS, EA 4135, F-33000, Bordeaux, France, Bordeaux INP, BPRVS, EA 4135, F-33000, Bordeaux, France.
authors: Bertrand
authors: B
authors: Garbay
authors: Univ Bordeaux, BPRVS, EA 4135, F-33000, Bordeaux, France, Bordeaux INP, BPRVS, EA 4135, F-33000, Bordeaux, France.
authors: Wilfrid
authors: W
authors: Dieryck
authors: Univ Bordeaux, Centre de Génomique Fonctionnelle, Plateforme Protéome, Bordeaux F-33000, France.
authors: Anne-Marie
authors: AM
authors: Lomenech
authors: Univ Bordeaux, BPRVS, EA 4135, F-33000, Bordeaux, France, Bordeaux INP, BPRVS, EA 4135, F-33000, Bordeaux, France.
authors: Xavier
authors: X
authors: Santarelli
authors: Univ Bordeaux, BPRVS, EA 4135, F-33000, Bordeaux, France, Bordeaux INP, BPRVS, EA 4135, F-33000, Bordeaux, France. Electronic address: charlotte.cabanne@bordeaux-inp.fr.
authors: Charlotte
authors: C
authors: Cabanne
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D016466:CHO Cells; D002846:Chromatography, Affinity / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods; D006224:Cricetinae; D003412:Cricetulus; D004247:DNA / Q000302:isolation & purification; D006801:Humans; D007074:Immunoglobulin G / Q000302:isolation & purification; D013058:Mass Spectrometry; D011506:Proteins / Q000737:chemistry; D011994:Recombinant Proteins / Q000302:isolation & purification; D013205:Staphylococcal Protein A / Q000737:chemistry
chemical_list: D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D011506:Proteins; D011994:Recombinant Proteins; D013205:Staphylococcal Protein A; D004247:DNA
keywords: Antibody; Host cell proteins; Mixed mode chromatography; Purification process