Potential Cytokines Enrichment Using Albusorb™ or Albuvoid™ for Albumin Depletion.

Potential Cytokines Enrichment Using Albusorb™ or Albuvoid™ for Albumin Depletion.

Albumin represents >90% protein in serum or plasma and it masks low-abundance proteins such as cytokines in sample preparation research. Cytokines represent important components of the inflammatory response in patients with trauma, shock, and sepsis. Humoral(interleukins 4, interleukin 5) reactions, immune reaction suppression (IL-10), cell differentiation (IL-2) and inflammatory responses (IFN-y, TNF-alpha) are mediated by cytokines. Pro-inflammatory cytokines worsen disease whereas other cytokines serve to reduce inflammation and enhance healing with beneficial anti-inflammatory effects on the body.

For the enrichment and isolation of cytokines, Biotech Support Group recommends implementing the Albusorb™ or AlbuVoid™ protocol for albumin depletion (removal of high abundance albumin) and/or potential low abundance cytokines/protein enrichment. AlbuVoid™ is unique because AlbuVoid™ does not bind albumin. Albusorb™ binds to albumin and other proteins are obtained in the flowthrough. Published articles on Albusorb™ explaining how Albusorb™ is used for albumin depletion:

• Effects of Nobiletin on PhIP-Induced Prostate and Colon Carcinogenesis in F344 Rats Authors: Ming Xi Tang, Nutrition and cancer. 02/2011; 63(2):227-33
  Authors showed how 0.05% citrus flavonoid nobiletin inhibited PhIP-induced rat prostate and colon carcinogenesis. Albusorb was used for albumin depletion from serum samples to enable leptin expression.
• Development of different analysis platforms with LC-MS for pharmacokinetic studies of protein drugs. Lu Q, Zheng X, Anal Chem. 2009 Nov 1;81(21):8715-23.
  Using Albusorb’s albumin depletion method first and then digest the depleted albumin solution (flow through fraction) for the subsequent LC-MS analysis of peptides, either 1-dimensional LC or 2-dimensional LC (ion exchange and reversed phase) with MS analysis.
• Apolipoprotein CIII and Ljungan virus in diabetes Holmberg, Rebecka Doctoral Thesis 26-May-2010 Albusorb was used on isolated islet cells from sera in prediabetic rats undergoing antisense treatment. Albusorb removed >90% albumin from serum samples.

Sample preparation steps involving depletion of high abundance proteins such as albumin is advantageous for low abundance cytokine discovery in serum or plasma. Scientists are increasingly focused on understanding why blocking of harmful cytokines during infection could solve diseases.

Subsequent to using Albusorb™ or AlbuVoid™, albumin depletion is measured using enzyme-linked immunoassay (ELISA), SDS-PAGE and cytokine recovery is determined by micro assay immunoassay that measures both pro- and anti- inflammatory cytokines.

Currently, Biotech Support Group is seeking research collaboration opportunities for studying how AlbuSorb™ or AlbuVoid™ affects albumin depletion and low abundance enrichment of proteins and cytokines.
For more information contact: sales@biotechsupportgroup.com

For more information:

• AlbuSorb™ Albumin Depletion Kit
• AlbuVoid™ - Albumin Depletion Kit from Serum or Plasma

Suggested References:

• Andrea Mahn, Alejandro Reyes, Mauricio Zamorano, Wildo Cifuentes, Maritza Ismail. (2010) Depletion of highly abundant proteins in blood plasma by hydrophobic interaction chromatography for proteomic analysis. Journal of Chromatography B 878:15-16, 1038-1044
• Muriel De Bock, Dominique de Seny, Marie-Alice Meuwis, Jean-Paul Chapelle, Edouard Louis, Michel Malaise, Marie- Paule Merville, Marianne Fillet. (2010) Challenges for Biomarker Discovery in Body Fluids Using SELDI-TOF-MS. Journal of Biomedicine and Biotechnology 2010, 1
• Assenmacher, M., Löhning, M., Scheffold, A., Manz, R. A., Schmitz, J., and Radbruch, A. 1998. Sequential production of IL-2,IFN-γ and IL-10 by individual staphylococcal enterotoxin B-activated T helper lymphocytes. Eur. J. Immunol. 28:1534- 1543.
• Granger, J., Siddiqui, J., Copeland, S. and Remick, D. (2005), Albumin depletion of human plasma also removes low abundance proteins including the cytokines. PROTEOMICS, 5: 4713–4718.
• Farrar, J.D., Ouyang, W., Löhning, M., Assenmacher, M., Radbruch, A., Kanagawa, O., and Murphy, K.M. 2001. An instructive component in T helper cell type 2 (Th2) development mediated by GATA-3. J. Exp. Med. 193:643-648.
• Hu-Li, J., Pannetier, C., Guo, L., Löhning, M., Gu, H., Watson, C., Assenmacher, M., Radbruch, A., and Paul, W.E. 2001. Regulation of expression of IL-4 alleles: Analysis using a chimeric GFP/IL-4 gene. Immunity 14:1-11.
• Manz, R., Assenmacher, M., Pfluger, E., Miltenyi, S., and Radbruch, A. 1995. Analysis and sorting of live cells according to secreted molecules relocated to a cell-surface affinity matrix. Proc. Natl. Acad. Sci. U.S.A. 92:1921-1925
• Ahmed N, Barker G, Oliva K, Garfin D, Talmadge K, Georgiou H, Quinn M, Rice G (2003) An approach to remove albumin for the proteomic analysis of low abundance biomarkers in human serum. Proteomics 3:1980–1987
• Adkins JN, Varnum SM, Auberry KJ, Moore RJ, Angell NH, Smith RD, Springer DL, Pounds JG (2002) Toward a human blood serum proteome. Analysis by multidimensional separation coupled with mass spectrometry. Mol Cell Proteomics 1:947– 955
• Anderson NL, Anderson NG (2002) The human plasma proteome: history, character, and diagnostic prospects. Mol Cell Proteomics 1:845–867
• Smits, H.H., van Rietschoten, J.G., Hilkens, C.M., Sayilir, R., Stiekema, F., Kapsenberg, M.L., and Wierenga, E.A. 2001. IL- 12-induced reversal of human Th2 cells is accompanied by full restoration of IL-12 responsiveness and loss of GATA-3 expression. Eur. J. Immunol. 31:1056-1065.
• Pittet, M.J., Zippelius, A., Speiser, D.E., Assenmacher, M., Guillaume, P., Valmori, D., Lienard, D., Lejeune, F., Cerottini, J.C., and Romero, P. 2001. Ex vivo IFN-gamma secretion by circulating CD8 T lymphocytes. Implications of a novel approach for T cell monitoring in infectious and malignant diseases. J. Immunol. 166:7634-7640.
• Oelke, M., Kurokawa, T., Hentrich, I., Behringer, D., Cerundolo, V., Lindemann, A., and Mackensen, A. 2000a. Functional characterization of CD8+ antigen-specific cytotoxic T lymphocytes after enrichment based on cytokine secretion: Comparison with the MHC-tetramer technology. Scand. J. Immunol. 52:544-549.
• Oelke, M., Moehrle, U., Chen, J.L., Behringer, D., Cerundolo, V., Lindemann, A., and Mackensen, A. 2000b. Generation and purification of CD8+ melan-A-specific cytotoxic T lymphocytes for adoptive transfer in tumor immunotherapy. Clin. Cancer Res. 6:1997-2005.
• Ouyang, W., Löhning, M., Gao, Z., Assenmacher, M., Ranganath, S., Radbruch, A., and Murphy, K.M. 2000. Stat6- independent GATA-3 autoactivation directs IL-4-independent Th2 development and commitment. Immunity 12:27-37.
• Brosterhus, H., Brings, S., Leyendeckers, H., Manz, R.A., Miltenyi, S., Radbruch, A., Assenmacher, M., and Schmitz, J. 1999. Enrichment and detection of live antigen-specific CD4+ and CD8+ T cells based on cytokine secretion. Eur. J. Immunol. 29:4053-4059.