Antibodies and Antibody Fragments

AN 3903 - MAb Purification from CCS with Pluronic using MAb A2P HF (08-01-2019)_V1

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Application Note MAbsorbent ® A2P HF Product Code: 3903 Application Note Purification of monoclonal antibodies from cell culture supernatant containing Pluronic ® F-68 using MAbsorbent ® A2P HF MAbsorbent ® A2P HF is a synthetic ligand affinity adsorbent designed and manufactured exclusively by Prometic Bioseparations Ltd (PBL). Our range of MAbsorbent ® products provide an alternative to Protein A, maintaining the requirements for binding capacity, yield and purity whilst operating in harsh environments for hundreds of process cycles. MAbsorbent ® A2P HF has a binding capacity in excess of 35 g/L for IgG and greater than 20 g/L for monoclonal antibody directly from cell culture supernatant (CCS) in the absence of Pluronic ® F-68. Pluronic ® F-68 is a linear polymer of isopropylene glycol repeating units and has properties similar to those of commonly used detergents such as Triton X-100. During the production of monoclonal antibodies, in mammalian cell culture supernatant, Pluronic ® F-68 is added to prevent sheer induced cell damage. However, at a concentration of 0.2 g/L, Pluronic ® F-68 significantly reduces the initial binding capacity of MAbsorbent ® A2P HF for monoclonal antibody directly from cell culture supernatant by approximately 60% and at a concentration of 0.5 g/L this is further reduced to greater than 90%. In this application note we describe the use of Amberchrom ™ CG161[1] for the removal of Pluronic ® F-68 from cell culture supernatant, containing monoclonal antibody, before the capture and purification of the antibody using MAbsorbent ® A2P HF. Pluronic ® F-68 structural image OH O O O y z x CH 3 H

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