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Nonionic detergent micelle aggregates: an economical alternative to Protein A chromatography
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  • Guy Patchornik,
  • Gunasekaran Dhandapani,
  • Ellen Wachtel,
  • Mordechai Sheves
Guy Patchornik
Ariel University Faculty of Natural Sciences
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Gunasekaran Dhandapani
Ariel University Faculty of Natural Sciences
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Ellen Wachtel
Weizmann Institute of Science Department of Biological Chemistry
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Mordechai Sheves
Weizmann Institute of Science Faculty of Chemistry
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Abstract

We have recently described a non-chromatographic, ligand-free approach for antibody (Ab) purification based on specially designed: [Tween-20:bathophenanthroline:Fe2+] aggregates. To assess the potential generality of this approach, a variety of detergents belonging to four nonionic detergent families (Tween, Brij, Triton and Pluronic) have now been studied. All surfactant aggregates lead to high purity of the recovered Abs (>95%, by gel densitometry). Good overall Ab recovery yields were observed with: Tween-20 (80-83%), Brij-O20 (85-87%) and Triton X-100 (87-90%), while Pluronic F-127 was less efficient (42-53%). Of additional importance is the finding that the process can depend on filtration (rather than centrifugation), thereby allowing a continuous purification mode that leads to the recovery of monomeric IgG’s (by DLS) and preservation of Ab specificity (by ELISA). The amphiphilic chelator, bathophenanthroline (batho) is recycled almost quantitatively (95%) by crystallization. Good IgG recovery yields (~80%) are also observed when Ab concentrations are increased from 1 mg/ml to 3-5 mg/mL. Potential advantages of the purification platform for industrial downstream processing of therapeutic monoclonal antibodies (mAbs), are discussed.

Peer review status:UNDER REVIEW

18 Feb 2020Submitted to Biotechnology and Bioengineering
18 Feb 2020Assigned to Editor
18 Feb 2020Submission Checks Completed
21 Apr 2020Reviewer(s) Assigned
26 May 2020Review(s) Completed, Editorial Evaluation Pending