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- Title
LYTAG‐driven purification strategies for monoclonal antibodies using quaternary amine ligands as affinity matrices.
- Authors
Campos‐Pinto, Isabel; Capela, Emanuel V.; Silva‐Santos, A. Rita; Rodríguez, Miguel Arévalo; Gavara, Poondi Rajesh; Fernandez‐Lahore, Marcelo; Aires‐Barros, M. Raquel; Azevedo, Ana M.
- Abstract
Abstract: BACKGROUND: Monoclonal antibodies are becoming a leading class of biopharmaceuticals but to increase their accessibility by the general population, new production processes must be developed in particular for the downstream processing. RESULTS: In this work, an alternative and innovative affinity chromatographic method using quaternary amine matrices is proposed. Separation is driven by the dual affinity ligand LYTAG‐Z, composed of a choline binding polypeptide (LYTAG) and the synthetic antibody binding Z domain. A two‐elution method was developed for the purification of mAbs and the performance of different anion exchangers containing quaternary amines that act as choline analogues – CIMmultus Q, Q Sepharose and gPore Q – were tested and compared, with both CIMmultus Q and Q Sepharose allowing a recovery of more than 94% of mAbs from a CHO cell supernatant with a purity greater than 95%. An integrated platform combining an initial affinity extraction step for the clarification and capture of mAbs and a subsequent chromatographic separation using Q‐matrices for the polishing of mAbs is also proposed. LYTAG‐Z triggers the extraction of 94.7 ± 1.7% mAbs to the PEG‐rich phase, as opposed to 26.9 ± 0.6% in the absence of the ligand, using 7% PEG 3350 and 6% dextran 500 k. Further purification using Q Sepharose allowed a mAb recovery of 95.3 ± 1.4% with a purity level of 91.4 ± 13.0%. CONCLUSION: An integrated platform based on two purification steps – affinity extraction and affinity chromatography – results in an overall process yield of 90%, allowing the processing of mAbs directly from a non‐clarified CHO cell culture. © 2017 Society of Chemical Industry
- Subjects
MONOCLONAL antibodies; LIGANDS (Chemistry); AMINES; BIOPHARMACEUTICS; AFFINITY chromatography; POLYPEPTIDES
- Publication
Journal of Chemical Technology & Biotechnology, 2018, Vol 93, Issue 7, p1966
- ISSN
0268-2575
- Publication type
Article
- DOI
10.1002/jctb.5460