IgG and IgM Purification and Quantitation
Monoclonal antibodies are one of the most important and fastest growing biological drugs. IgG's are being purified using established DSP protocols and produced in kilogram scale. Recently, immunoglobulins M (IgM's) have become increasingly important as they have a different selectivity and can cope with certain diseases in a more efficient way.
IgM's are extremely large proteins (up to 1 M Da). As such they are more susceptible to shear forces that may occur during purification on chromatography columns. As well, they are less stable in lower pH or higher ionic-strength environment and prone to form aggregates. Therefore, the purification process that would completely overcome the above mentioned problems is still a challenge.
CIM® Monolithic columns can overcome some of these challenges. They have the open channel structure where large molecules can freely and openly be conveyed without any steric hindrances. Also, the contact time between the molecules and the stationary phase can be significantly reduced (a combination of a short separation layer and high volumetric flow rates applicable), thus substantially reducing the risks of negatively impacting the labile IgM molecules during purification process.
See how these features reflect in the applications presented below.
| A024 | Rapid Separation of IgG, IgA and IgM on a CIM® QA Disk Monolithic Column |
| A009 | Quantitative Determination of Human IgG by High-Performance Immunoaffinity Chromatography Using CIM® Protein A Disk |
| A019 | Separation of IgG, HSA and IgM Standard on CIM® EDA Monolithic Column |
| A010 | Isolation of Anti-Bradikinin IgGs from Crude Serum of Immunized Rabbit by High-Performance Immunoaffinity Chromatography Using CIM® Bradikinin Disk |
| P003 | Capacity Determination for Binding Human IgG to a CIM® SO3-8 Tube Monolithic Column For additional information please see the following: Poster |
| P004 | Productivity of Semi-Preparative Isolation of Immunoglobulins G from Human Plasma on CIM® Disk and CIM® Tube Protein G Monolithic Columns Versus Particle Based Agarose Supports For additional information please see the following: Poster |
| A004 | Conjoint Liquid Chromatography (CLC) Separation of Proteins from Mouse Ascites and Isolation of Monoclonal Antibody IgG in One Step Obtained by a Combination of CIM® QA and CIM® Protein A Disks |
Available posters
More information on IgG and IgM purification/quantitation on CIM Monoliths can be found at Validated Biosystems Inc. website (courtesy of P. Gagnon).
- A high speed monolithic assay for IgM quantitation in cell culture production and purification process monitoring
- A comparison of microparticulate, membrane, and monolithic anion exchangers for polishing applications in the purification of IgG monoclonal antibodies
Peer-reviewed articles
See below a list of articles on Monolith IgG and IgM purification that are available in various on-line libraries such as ScienceDirect. Article abstracts are also available through our website's search engine - here.
- Chromatographic behavior of IgM:DNA complexes, P. Gagnon, F. Hensel, S. Lee, S. Zaidi - Journal of Chromatography A, 1218 (2011) 2405-2412
- Dissociation and fractionation of heavy and light chains from IgG monoclonal, P. Gagnon, G. Rodriquez, S. Zaidi - Journal of Chromatography A, 1218 (2011) 2402-2404
- Monolith peptide affinity chromatography for quantification of immunoglobulin M, S. Neff, A. Jungbauer - Journal of Chromatography A, 1218 (2011) 2374-2380
- Purification of Monoclonal antibodies from cell-culture supernatant by use of anion-exchange Convective Interaction Media (CIM) Monolithic Columns, A. Dhivya, B. Kumar, R. Prasanna, N. Vijayalakshmi - Chromatographia 2010, 72, December (No. 11/12), pg 1183-1188
- Characterization of Metal Chelate Methacrylate Monolithic Disk for Purification of Polyclonal and Monoclonal Immunoglobulin G, R. R. Prasanna - Journal of Chromatography A, 1217 (2010) 3660–3667
- Engineering of a Two-Step Purification Strategy for a Panel of Monoclonal Immunoglobulin M Directed Against Undifferentiated Human Embryonic Stem Cells, A. Tscheliessnig, D. Ong, J. Lee, S. Pan, G. Satianegara, K. Schriebl, A. Choo, A. Jungbauer - Journal of Chromatography A, 1216 (2009) 7851–7864
- Application of Conjoint Liquid Chromatography with Monolithic Disks for the Simultaneous Determination of Immunoglobulin G and Other Proteins Present in a Cell Culture Medium, K. Ralla, F. Anton, T. Scheper, C. Kasper - Journal of Chromatography A, 1216 (2009) 2671–2675
- High-Performance Monolith Affinity Chromatography for Fast Quantitation of Immunoglobulin G, A. Tscheliessnig, A. Jungbauer - ( Journal of Chromatography A, 1216 (2009) 2676–2682
- Fast and Efficient Separation of Immunoglobulin M from Immunoglobulin G using Short Monolithic Columns, P. Brne, A. Podgornik, K. Bencina, B. Gabor, A. Strancar, M. Peterka - Journal of Chromatography A, 1144 (2007) 120–125
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