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Purification of monoclonal antibodies, IgG1, from cell culture supernatant by use of metal chelate convective interaction media monolithic columns
Rajak P, Vijayalakshmi M.A,
Published in Wiley
PMID: 22362585
Volume: 26
Issue: 12
Pages: 1488 - 1493
Monoclonal antibodies (MAbs) have diverse applications in diagnostics and therapeutics. The recent advancement in hybridoma technology for large-scale production of MAbs in bioreactors demands rapid and efficient purification methods. Conventional affinity purification systems have drawbacks of low flow rates and denaturation of antibodies owing to harsh elution conditions. Here, we attempted purification of MAbs by use of a high-throughput metal-chelate methacrylate monolithic system. Monolithic macroporous convective interaction media-iminodiacetate (CIM-IDA) disks immobilized with four different metal ions (Cu2+, Ni2+, Zn2+ and Co2+) were used and evaluated for purification of anti-human serum albumin IgG1 mouse MAbs from cell culture supernatant after precipitation with 50% ammonium sulfate. Elution with 10mM imidazole in the equilibration buffer (25mM MMA=MOPS (Morpholino propane sulfonic acid)+MES (Morpholino ethane sulfonic acid)+Acetate+0.5M NaCl, pH 7.4) resulted in a purification of 25.7 ± 2.9-fold and 32.5 ± 2.6-fold in experiments done using Zn2+ and Co2+ metal ions, respectively. The highest recovery of 85.4 ± 1.0% was obtained with a CIM-IDA-Zn(II) column. SDS-PAGE, ELISA and immuno-blot showed that the antibodies recovered were pure, with high antigen-binding efficiency. Thus, metal chelate CIM monoliths could be a potential alternative to conventional systems for fast and efficient purification of MAbs from the complex cell culture supernatant. © 2012 John Wiley & Sons, Ltd.
About the journal
JournalData powered by TypesetBiomedical Chromatography
PublisherData powered by TypesetWiley
Open Access0