Ascites fluid and cell culture clarification via Cleanascite™ for monoclonal antibody purification

PRLog (Press Release) - Aug. 12, 2012 - MONMOUTH JUNCTION, NJ.– Cleanascite™  from Biotech Support Group cleans up samples and it extends the life of membrane and chromatographic columns for large scale monoclonal antibody purification. Before purifying hybridoma cell cultures or ascites, clarification is required. Cleanascite™ selectively removes lipids, cell debris, lipoproteins, floating fats, impurities from cohn paste, transgenic milk, egg yolk and biological samples for pretreatment of samples prior to purification. Cleanascite™ is used for clarifying samples either from cell culture or ascites during monoclonal antibody purification. Usually hybridoma cells are immobilized in sol–gel silica matrices for in vitro production of monoclonal antibodies. Hybridoma cell lines that do not grow or produce antibody by in vitro methods require in vivo ascites method. In vivo methods to produce monoclonal antibodies have approved protocols which grow hybridoma cells within the peritoneal cavity of a mouse.  Ascites is produced upon injection of hybridoma cells in a mouse. The ascites fluid is contaminated with bioreactive cytokines, rodent plasma proteins, infectious agents and immunoglobulins. It also has a high concentration of antibody.

Cleanascite™ is ideal for bioprocessing while producing therapeutic monoclonal antibodies (mAbs). The reagent is a solid-phase, non-ionic adsorbent supplied as a suspension in saline, ready for use. Simply add, centrifuge and/or filter. The clarified supernatant is ready for subsequent downstream processing or analysis. It helps purify antibodies, recombinant proteins, nucleic acids, proteoglycans. It is also ideal for clarifying ascites, serum, cell & tissue culture, bile and organ homogenates.

Application of Cleanascite™ for clarification of ascites during monoclonal antibody production:
Nussbaum G, Cleare W, Casadevall A et al. Epitope Location in the Cryptococcus neoformans Capsule Is a Determinant of Antibody Efficacy. The Journal of experimental medicine.1997;185:685-694

For the preparation of monoclonal antibodies from ascites of hybridoma cells, Cleanascite™ was used for lipid removal. Monoclonal antibodies (mAbs) to the polysaccharide capsule of Cryptococcus neoformans can prolong survival in mice. epitope specificity in determining protective efficacy was suggested by experiments with two murine IgM anticryptococcal mAbs, 12A1 and 13F1.  The protective mAb, 12A1, produced a homogeneous annular fluorescence pattern, whereas the nonprotective mAb, 13F1, produced a punctate pattern of fluorescence on one strain of serotype D, C. neoformans.

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http://www.biotechsupportgroup.com/node/73

About Biotech Support Group LLC
Biotech Support Group LLC is a leading provider of genomics and proteomics sample preparation products and enrichment reagent kits as well as integrated biotechnology services for life sciences research, biomarker and drug discovery. Based in New Jersey, it’s principal products include:  AlbuVoid™ for albumin depletion, Cleanascite™ for lipid adsorption and clarification, NuGel™ for passivated silica-based affinity chromatography, and ProCipitate™ & ProPrep™ for nucleic acid isolation. Currently, Biotech Support Group LLC and ProFACT Proteomics Inc., are collaborating on the development of a proteomics platform used in functional profiling for proteomic analysis and a separations method for generating sub-proteomes used in biomarker and functional proteomic prospecting. For more information, go to: www.biotechsupportgroup.com.

CONTACT:
Dr.Swapan Roy
Biotech Support Group            
1 Deer Park Drive, Suite M,
Monmouth Junction, NJ 08852, USA              
732-274-2866              
sales@biotechsupportgroup.com

References for Cleanascite™
1.J Krupey - United States Patent: 5885921 Hydrophobic silica adsorbents for lipids.
2.Cho N, Chueh PJ, Kim C et al Monoclonal antibody to a cancer-specific and drug-responsive hydroquinone (NADH) oxidase from the sera of cancer patients. Cancer Immunology, Immunotherapy. 2002;51(3):121-9
3.Hauser-Davis RA, Lima AA, Ziolli RL, Campos RC.First-time report of metalloproteinases in fish bile and their potential as bioindicators regarding environmental contamination. Aquatic Toxicology.2012;110-111:99-106
4.Beenhouwer DO, Shapiro S, Feldmesser M et al. Both Th1 and Th2 Cytokines Affect the Ability of Monoclonal Antibodies To Protect Mice against Cryptococcus neoformansInfection and immunity.2001;69: 6445-6455
5.Nussbaum G, Cleare W, Casadevall A et al Epitope Location in the Cryptococcus neoformans Capsule Is a Determinant of Antibody Efficacy The Journal of experimental medicine.1997;185:685-694

Suggested References
1.Kamp, M. and W. de Leeuw. Short review of in vitro production methods for monoclonal antibodies. NCA Newsletter.1996;3:10-11
2.Falkenberg, F. W., Hengelage, T., Krane, M., Bartels, I., Albrecht, A., Holtmeier, N., and M. Wuthrich. A simple and inexpensive high density dialysis tubing cell culture system for the in vitro production of monoclonal antibodies in high concentrations. Journal of Immunological Methods.1993; 165:193-206
3.Bodeus, M., Bartonboy, G., and H. Bazin. Rat monoclonal antibodies IV. Easy method for in vitro production. Journal of Immunological Methods.1985; 79:1-6   
4.Sjorgren-Jansson, E. and S. Jeansson (1990). Growing hybridomas in dialysis tubing, optimization of the technique. In Laboratory Methods Immunology. 1990. CRC Press: Boca Raton, FL, pp. 1-41
5.McArdle, J., Reddington, J., Reddington, G., and J. Heidel (1996). Further development of a unique, simple, inexpensive, efficient in vitro method for small, medium and large scale production of monoclonal antibodies.1996; ATLA 24 (special issue):318
6.Monica, T.J., et al. Comparative biochemical characterization of a human IgM produced in both ascites and in vitro cell culture.1993; BioTechnology11:512-515.
7.Peterson, N.C. Considerations for in vitro monoclonal antibody production. Research In Immunology. 1998; 149(6):553-557.

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