Advances in Technology and Process Development for Industrial‐Scale Monoclonal Antibody Purification

Fontes, Nuno; Reis, Robert van

doi:10.1002/9780470444894.ch10

Cited by 5 publications

(1 citation statement)

References 21 publications

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“…Purification of monoclonal antibodies from crude cell culture feedstocks thus requires multi-step processes, which have in the past included membrane-based tangential flow filtration methods, aqueous two phase separation procedures and affinity, ion exchange and hydrophobic interaction chromatography [6][7][8][9][10][11][12]. Frequently, affinity chromatography has been employed as the preferred separation method at the laboratory or process scale [13] with the traditional ligands based on bacterial Fc receptors, such as Protein A from Staphylococcus aureus, or Protein G from Streptococcus sp, favoured [5,14,15].…”

Section: Introductionmentioning

confidence: 99%

Purification of monoclonal antibodies by chemical affinity mixed mode chromatography

Zhang

Fredericks

Campi

et al. 2015

Separation and Purification Technology

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Section: Introductionmentioning

confidence: 99%

Purification of monoclonal antibodies by chemical affinity mixed mode chromatography

Zhang

Fredericks

Campi

et al. 2015

Separation and Purification Technology

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Dual salt mixtures in mixed mode chromatography with an immobilized tryptophan ligand influence the removal of aggregated monoclonal antibodies

Vajda¹,

Mueller²,

Bahret

2014

Biotechnology Journal

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In downstream processing of monoclonal antibodies, proper aggregate removal is crucial. Mixed mode ligands such as immobilized tryptophan have been developed to satisfy the need for efficient removal of antibody aggregates. However, method development for mixed mode applications is complicated, since protein binding and elution can be modulated by an increased set of parameters. In the current study, we investigate the effect of different dual salt mixtures on mixed mode chromatography using TOYOPEARL MX-Trp-650M resin, with respect to the dynamic binding capacity, resolution and monomer purity of two different humanized immunoglobulins. Binding capacities varying by more than 50% were observed for different salt mixtures. Furthermore, antibody monomer and aggregate resolution deviated by 30% for different salt mixtures and linear gradient elution. Similar trends were obtained using an immobilized carboxymethyl ligand for the same set of experiments, but the overall resolution was lower. Less kosmotropic salt systems emphasize the electrostatic binding of the relatively hydrophobic mAbs and reduce hydrophobic attraction to a selectivity-determining constraint. Kosmotropic salts such as citrate appear to cause dominating hydrophobic interactions in protein adsorption that hinder electrostatic protein-ligand interactions. This effect may depend on the ionic and hydrophobic site distribution of a protein. The data presented here are important for the further improvement of downstream processing of therapeutic monoclonal antibodies.

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Increasing parvovirus filter throughput of monoclonal antibodies using ion exchange membrane adsorptive pre‐filtration

Brown

Bechtel

Bill

et al. 2010

Biotech & Bioengineering

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Pre-filtration using ion exchange membrane adsorbers can improve parvovirus filter throughput of monoclonal antibodies (mAbs). The membranes work by binding trace foulants, and although some antibody product also binds, yields > or =99% are easily achieved by overloading. Results show that foulant adsorption is dependent on pH and conductivity, but independent of scale and adsorber brand. The ability to use ion exchange membranes as pre-filters is significant because it provides a clean, well defined, chemically stable option for enhancing throughput. Additionally, ion exchange membranes facilitate characterization of parvovirus filter foulants. Examination of adsorber elution samples using sedimentation velocity analysis and SEC-MALS/QELS revealed the presence of high molecular weight species ranging from 8 to 13 nm in hydrodynamic radius, which are similar in size to parvoviruses and thus would be expected to plug the pores of a parvovirus filter. A study of two identical membranes in-series supports the hypothesis that the foulants are soluble, trace level aggregates in the feed. This study's significance lies in a previously undiscovered application of membrane chromatography, leading to a more cost effective and robust approach to parvovirus filtration for the production of monoclonal antibodies.

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Advances in Technology and Process Development for Industrial‐Scale Monoclonal Antibody Purification

Cited by 5 publications

References 21 publications

Purification of monoclonal antibodies by chemical affinity mixed mode chromatography

Purification of monoclonal antibodies by chemical affinity mixed mode chromatography

Dual salt mixtures in mixed mode chromatography with an immobilized tryptophan ligand influence the removal of aggregated monoclonal antibodies

Increasing parvovirus filter throughput of monoclonal antibodies using ion exchange membrane adsorptive pre‐filtration

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