Monolithic convective interaction media-disk (CIM-disk) chromatography is one of the fastest liquid chromatographic methods for the separation and purification of biomolecules due to its high mass transfer rate. In this way, all separated molecules are transported by convection into the pores of the matrix, resulting in a very fast separation due to the low mass transfer resistance of the CIM-disk. Due to the advantages of monolith technique, in recent years, CIM-disk affinity chromatography has been developed and investigated for purification of peptides, restriction enzymes, antibodies, etc. In this review, applications of monolithic affinity chromatography are discussed. The purification of restriction enzymes, polyclonal and monoclonal antibodies using a new monolithic CIM-disk system with immobilized histidine affinity chromatography are presented. © 2007 Friedr. Vieweg &amp; Sohn Verlag/GWV Fachverlage GmbH.

Shanthi C

Department of Biotechnology

School of Bio Sciences and Technology

Vellore Campus

Satheesh A

Department of Thermal and Energy Engineering

School of Mechanical Engineering

Champagne J

Delattre C

Duverneuil L

Vijayalakshmi M.A.

Vellore Institute of Technology (VIT) is a private university located in&nbsp;Tamil Nadu, India. Founded in 1984, as Vellore Engineering College, the institution offers 20 undergraduate, 34 postgraduate, four integrated and four research programs. It has campuses in Vellore, Amravati, Bhopal and Chennai.

VIT is one of the top ranked private universities in India according to NIRF, THE and QS Rankings.&nbsp;Govt. of India has recognized&nbsp;VIT, Vellore as an&nbsp;Institution of Eminence. This has allowed VIT to take independent quality initiatives and move up in world ranking.

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VIT University

Chromatographia

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 &amp; Sons, Ltd.

Biomedical Chromatography

Purification of monoclonal antibodies, IgG1, from cell culture supernatant by use of metal chelate convective interaction media monolithic columns

Purified monoclonal antibodies (mAb) have been used in therapeutics and some analytical procedures. Purification of mAb by use of high-throughput anion-exchange methacrylate monolithic systems has been attempted in this work. Monolithic macroporous convective interaction media (CIM) with diethylaminoethyl (DEAE) and ethylene diamine (EDA) as anion-exchange ligands were used and evaluated for purification of anti-glycophorin-A IgG1 mouse mAbs from cell culture supernatant (CCS) after precipitation with 50% ammonium sulfate. The adsorption and elution of mAb from the CCS on CIM-DEAE and CIM-EDA disks were studied with three different buffer systems, acetate, MOPS (3-(N-morpholino) propanesulfonic acid), and Tris, to study the effect of the nature of buffer ions and to find the optimum buffer conditions for purification of mAb. The optimum buffers for purification of mAb using CIM-DEAE and CIM-EDA were 50 mM acetate buffer, pH 5.1 and 20 mM Tris buffer, pH 8.0, respectively. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and enzyme-linked immunosorbent assay (ELISA) showed the antibody fractions obtained were highly pure, with high antigen-binding efficiency. High specific activity with purification factors of 130 ± 34 (unretained fraction with acetate buffer) and 74 ± 13 (fraction eluted with Tris buffer containing 0.6 M NaCl) was obtained for IgG1 using the CIM-DEAE and CIM-EDA disks, respectively. The results indicate that rapid separation and efficient recovery of high-purity anti-glycophorin-A mAbs could be achieved by use of anion-exchange CIM disks. © 2010 Vieweg+Teubner Verlag | Springer Fachmedien Wiesbaden GmbH.

Purification of Monoclonal Antibodies from Cell-Culture Supernatant by Use of Anion-Exchange Convective Interaction Media (CIM) Monolithic Columns

Journal of Molecular Catalysis B: Enzymatic

Monolith enzymatic microreactor at the frontier of glycomic toward a new route for the production of bioactive oligosaccharides

Biotechnology Journal

Downstream processing of value-added proteins in biotechnology: An Indian scenario

10.1002/bmc.v29.1

ELECTROPHORESIS

Affinity monolithic capillary columns for glycomics/proteomics: 1. Polymethacrylate monoliths with immobilized lectins for glycoprotein separation by affinity capillary electrochromatography and affinity nano-liquid chromatography in either a single column or columns coupled in series

Journal of Proteome Research

Proteomic Analysis of Serum Associated Fucosylated Glycoproteins in the Development of Primary Hepatocellular Carcinoma

Journal of Chromatography A

Purification of oligouronides using hollow-fiber membrane functionalised with l-histidine

Journal of Separation Science

Convective Interaction Media short monolithic columns: Enabling chromatographic supports for the separation and purification of large biomolecules

Pseudoaffinity Chromatography Using a Convective Interaction Media®-Disk Monolithic Column

Journal	Data powered by TypesetChromatographia
Publisher	Data powered by TypesetSpringer Science and Business Media LLC
ISSN	0009-5893
Open Access	0